Clinical impact of CEUS on non-characterizable observations and observations with intermediate probability of malignancy on CT/MRI in patients at risk for HCC.

BACKGROUND: Hepatocellular carcinoma (HCC) is a unique cancer allowing tumor diagnosis with identification of definitive patterns of enhancement on contrast-enhanced imaging, avoiding invasive biopsy. However, it is still unclear to what extent Contrast-Enhanced Ultrasound (CEUS) is a clinically useful additional step when Computed tomography (CT) or Magnetic resonance imaging (MRI) are inconclusive. METHODS: A prospective international multicenter validation study for CEUS Liver Imaging Reporting and Data System (LI-RADS) was conducted between January 2018 and August 2021. 646 patients at risk for HCC with focal liver lesions were enrolled. CEUS was performed using an intravenous ultrasound contrast agent within 4 weeks of CT/MRI. Liver nodules were categorized based on LI-RADS (LR) criteria. Histology or one-year follow-up CT/MRI imaging results were used as the reference standard. The diagnostic performance of CEUS was evaluated for inconclusive CT/MRI scan in two scenarios for which the AASLD recommends repeat imaging or imaging follow-up: observations deemed non-characterizable (LR-NC) or with indeterminate probability of malignancy (LR-3). RESULTS: 75 observations on CT or MRI were categorized as LR-3 (n = 54) or LR-NC (n = 21) CEUS recategorization of such observations into a different LR category (namely, into one among LR-1, LR-2, LR-5, LR-M, or LR-TIV) resulted in management recommendation changes in 33.3% (25/75) and in all but one (96.0%, 24/25) observation, the new management recommendations were correct. CONCLUSION: CEUS LI-RADS resulted in management recommendations change in substantial number of liver observations with initial indeterminate CT/MRI characterization, identifying both non-malignant lesions and HCC, potentially accelerating the diagnostic process and alleviating the need for biopsy or follow-up imaging. CLINICALTRIALS: gov number, NCT03318380.

Transrectal Subharmonic Ultrasound Imaging for Prostate Cancer Detection.

OBJECTIVE: To assess the prostate cancer (CaP) detection rates of contrast-enhanced, transrectal subharmonic ultrasound imaging (SHI). MATERIALS AND METHODS: This IRB-approved study enrolled 55 subjects. The initial 5 subjects were studied for SHI optimization, while the remaining 50 were evaluated with contrast-enhanced sonography using continuous SHI, color, and power Doppler as well as conventional grayscale, continuous color, and power Doppler and SHI combined with maximum flash replenishment. A maximum of 6 directed biopsy cores were obtained from sites of greatest asymmetrical enhancement, followed by spatially distributed cores in a double sextant distribution. Subharmonic time-intensity parameters, including time to peak intensity, peak intensity, and estimated perfusion were also evaluated for each directed biopsy core. Receiver operating characteristic curve analysis and conditional logistic regression were employed to assess the benefit of each modality and the quantitative SHI parameters. RESULTS: Cancer was detected in 22 of 50 subjects. Among subjects with clinically significant CaP (n = 11), targeted cores were more likely to be positive (odds ratio 1.39, P = .02). The majority of patients detected by SHI demonstrated significant CaP (5/8); SHI remained an independent marker of malignancy in a multivariate logistic regression model (P = .027). Receiver operating characteristic curve analysis of imaging findings compared to biopsy results yielded diagnostic accuracies ranging from 0.59 to 0.80 for all imaging modalities with the highest being for quantitative subharmonic perfusion estimates. CONCLUSION: This first-in-humans study provides a preliminary estimate of the diagnostic accuracy of SHI for detection of clinically significant CaP (up to 80%).

Subharmonic aided pressure estimation for monitoring interstitial fluid pressure in tumours--in vitro and in vivo proof of concept.

The feasibility of using subharmonic aided pressure estimation (SHAPE) to noninvasively estimate interstitial fluid pressure (IFP) was studied. In vitro, radiofrequency signals, from 0.2 ml/l of Definity (Lantheus Medical Imaging, N Billerica, MA) were acquired within a water-tank with a Sonix RP ultrasound scanner (Analogic Ultrasound, Richmond, BC, Canada; fT/R=6.7/3.35 MHz and fT/R=10/5 MHz) and the subharmonic amplitudes of the signals were compared over 0-50 mmHg. In vivo, five swine with naturally occurring melanomas were studied. Subharmonic signals were acquired from tumours and surrounding tissue during infusion of Definity and compared to needle-based pressure measurements. Both in vitro and in vivo, an inverse linear relationship between hydrostatic pressure and subharmonic amplitude was observed with r(2)=0.63-0.95; p<0.05, maximum amplitude drop 11.36 dB at 10 MHz and -8 dB, and r(2) as high as 0.97; p<0.02 (10 MHz and -4/-8 dB most promising), respectively, indicating that SHAPE may be useful in monitoring IFP.

Electromechanical piezoresistive sensing in suspended graphene membranes.

Monolayer graphene exhibits exceptional electronic and mechanical properties, making it a very promising material for nanoelectromechanical devices. Here, we conclusively demonstrate the piezoresistive effect in graphene in a nanoelectromechanical membrane configuration that provides direct electrical readout of pressure to strain transduction. This makes it highly relevant for an important class of nanoelectromechanical system (NEMS) transducers. This demonstration is consistent with our simulations and previously reported gauge factors and simulation values. The membrane in our experiment acts as a strain gauge independent of crystallographic orientation and allows for aggressive size scalability. When compared with conventional pressure sensors, the sensors have orders of magnitude higher sensitivity per unit area.

On the implementation of an automated acoustic output optimization algorithm for subharmonic aided pressure estimation.

Incident acoustic output (IAO) dependent subharmonic signal amplitudes from ultrasound contrast agents can be categorized into occurrence, growth or saturation stages. Subharmonic aided pressure estimation (SHAPE) is a technique that utilizes growth stage subharmonic signal amplitudes for hydrostatic pressure estimation. In this study, we developed an automated IAO optimization algorithm to identify the IAO level eliciting growth stage subharmonic signals and also studied the effect of pulse length on SHAPE. This approach may help eliminate the problems of acquiring and analyzing the data offline at all IAO levels as was done in previous studies and thus, pave the way for real-time clinical pressure monitoring applications. The IAO optimization algorithm was implemented on a Logiq 9 (GE Healthcare, Milwaukee, WI) scanner interfaced with a computer. The optimization algorithm stepped the ultrasound scanner from 0% to 100% IAO. A logistic equation fitting function was applied with the criterion of minimum least squared error between the fitted subharmonic amplitudes and the measured subharmonic amplitudes as a function of the IAO levels and the optimum IAO level was chosen corresponding to the inflection point calculated from the fitted data. The efficacy of the optimum IAO level was investigated for in vivo SHAPE to monitor portal vein (PV) pressures in 5 canines and was compared with the performance of IAO levels, below and above the optimum IAO level, for 4, 8 and 16 transmit cycles. The canines received a continuous infusion of Sonazoid microbubbles (1.5 µl/kg/min; GE Healthcare, Oslo, Norway). PV pressures were obtained using a surgically introduced pressure catheter (Millar Instruments, Inc., Houston, TX) and were recorded before and after increasing PV pressures. The experiments showed that optimum IAO levels for SHAPE in the canines ranged from 6% to 40%. The best correlation between changes in PV pressures and in subharmonic amplitudes (r=-0.76; p=0.24), and between the absolute PV pressures and the subharmonic amplitudes (r=-0.89; p<0.01) were obtained for the optimized IAO and 4 transmit cycles. Only for the optimized IAO and 4 transmit cycles did the subharmonic amplitudes differ significantly (p<0.01) before and after increasing PV pressures. A new algorithm to identify optimum IAO levels for SHAPE has been developed and validated with the best results being obtained for 4 transmit cycles. The work presented in this study may pave the way for real-time clinical applications of estimating pressures using the subharmonic signals from ultrasound contrast agents.

Guidelines and good clinical practice recommendations for contrast enhanced ultrasound (CEUS) in the liver--update 2012: a WFUMB-EFSUMB initiative in cooperation with representatives of AFSUMB, AIUM, ASUM, FLAUS and ICUS.

Initially, a set of guidelines for the use of ultrasound contrast agents was published in 2004 dealing only with liver applications. A second edition of the guidelines in 2008 reflected changes in the available contrast agents and updated the guidelines for the liver, as well as implementing some non-liver applications. Time has moved on, and the need for international guidelines on the use of CEUS in the liver has become apparent. The present document describes the third iteration of recommendations for the hepatic use of contrast enhanced ultrasound (CEUS) using contrast specific imaging techniques. This joint WFUMB-EFSUMB initiative has implicated experts from major leading ultrasound societies worldwide. These liver CEUS guidelines are simultaneously published in the official journals of both organizing federations (i.e., Ultrasound in Medicine and Biology for WFUMB and Ultraschall in der Medizin/European Journal of Ultrasound for EFSUMB). These guidelines and recommendations provide general advice on the use of all currently clinically available ultrasound contrast agents (UCA). They are intended to create standard protocols for the use and administration of UCA in liver applications on an international basis and improve the management of patients worldwide.

Simultaneous grayscale and subharmonic ultrasound imaging on a modified commercial scanner.

OBJECTIVE: To demonstrate the feasibility of simultaneous dual fundamental grayscale and subharmonic imaging on a modified commercial scanner. MOTIVATION: The ability to generate signals at half the insonation frequency is exclusive to ultrasound contrast agents (UCA). Thus, subharmonic imaging (SHI; transmitting at f(0) and receiving at f(0)/2) provides improved visualization of UCA within the vasculature via suppression of the surrounding tissue echoes. While this capability has proven useful in a variety of clinical applications, the SHI suppression of surrounding tissue landmarks (which are needed for sonographic navigation) also limits it use as a primary imaging modality. In this paper we present results using a commercial ultrasound scanner modified to allow imaging in both grayscale (f(0)=4.0 MHz) and SHI (f(0)=2.5 MHz, f(0)/2=1.25 MHz) modes in real time. METHODS: A Logiq 9 ultrasound scanner (GE Healthcare, Milwaukee, WI) with a 4C curvilinear probe was modified to provide this capability. Four commercially available UCA (Definity, Lantheus Medical Imaging, North Billerica, MA; Optison, GE Healthcare, Princeton, NJ; SonoVue, Bracco Imaging, Milan, Italy; and Sonazoid, GE Healthcare, Oslo, Norway) were all investigated in vitro over an acoustic output range of 3.34 MPa. In vivo the subharmonic response of Sonazoid was investigated in the portal veins of four canines (open abdominal cavity) and four patients with suspected portal hypertension. RESULTS: In vitro, the four UCA showed an average maximum subharmonic amplitude of 44.1±5.4 dB above the noise floor with a maximum subharmonic amplitude of 48.6±1.6 dB provided by Sonazoid. The average in vivo maximum signal above the noise floor from Sonazoid was 20.8±2.3 dB in canines and 33.9±5.2 dB in humans. Subharmonic amplitude as a function of acoustic output in both groups matched the S-curve behavior of the agent observed in vitro. The dual grayscale imaging provided easier sonographic navigation, while the degree of tissue suppression in SHI mode varied greatly on a case by case basis. CONCLUSIONS: These results demonstrate the feasibility of dual grayscale and SHI on a modified commercial scanner. The ability to simultaneously visualize both imaging modes in real time should improve the applicability of SHI as a future primary clinical imaging modality.

Contrast-enhanced ultrasound for molecular imaging of angiogenesis.

INTRODUCTION: Molecular imaging of angiogenesis using contrast-enhanced ultrasound allows for functional, real-time, inexpensive imaging of angiogenesis. The addition of stabilized microbubbles as contrast agents greatly improves ultrasound signal to noise ratio/signal strength/image quality (up to 25 dB) and allows for imaging of angiogenic vasculature. METHODS: In this article recent advances in the usage of contrast-enhanced ultrasound for molecular imaging of angiogenesis are reviewed. RESULTS: The usage of commercially available agents and correlations between their imaging parameters and molecular markers of angiogenesis are reviewed. Recent developments in ultrasound contrast agents targeted to angiogenic markers for both diagnosis and monitoring are discussed. Finally, a brief overview of the emerging field of chemotherapeutic-loaded agents, which can be used with ultrasound-triggered drug delivery, is provided.

Development and optimization of a doxorubicin loaded poly(lactic acid) contrast agent for ultrasound directed drug delivery.

An echogenic, intravenous drug delivery platform is proposed in which an encapsulated chemotherapeutic can travel to a desired location and drug delivery can be triggered using external, focused ultrasound at the area of interest. Three methods of loading poly(lactic acid) (PLA) shelled ultrasound contrast agents (UCA) with doxorubicin are presented. Effects on encapsulation efficiency, in vitro enhancement, stability, particle size, morphology and release during UCA rupture are compared by loading method and drug concentration. An agent containing doxorubicin within the shell was selected as an ideal candidate for future hepatocellular carcinoma studies. The agent achieved a maximal drug load of 6.2 mg Dox/g PLA with an encapsulation efficiency of 20.5%, showed a smooth surface morphology and tight size distribution (poly dispersity index=0.309) with a peak size of 1865 nm. Acoustically, the agent provided 19 dB of enhancement in vitro at a dosage of 10 microg/ml, with a half life of over 15 min. In vivo, the agent provided ultrasound enhancement of 13.4+/-1.6 dB within the ascending aorta of New Zealand rabbits at a dose of 0.15 ml/kg. While the drug-incorporated agent is thought to be well suited for future drug delivery experiments, this study has shown that agent properties can be tailored for specific applications based on choice of drug loading method.

3D micro-scale deformations of wood in bending: synchrotron radiation muCT data analyzed with digital volume correlation.

A micro-scale three-point-bending experiment with a wood specimen was carried out and monitored by synchrotron radiation micro-computed tomography. The full three-dimensional wood structure of the 1.57x3.42x0.75mm(3) specimen was reconstructed at cellular level in different loading states. Furthermore, the full three-dimensional deformation field of the loaded wood specimen was determined by digital volume correlation, applied to the reconstructed data at successive loading states. Results from two selected regions within the wood specimen are presented as continuous displacement and strain fields in both 2D and 3D. The applied combination of synchrotron radiation micro-computed tomography and digital volume correlation for the deformation analysis of wood under bending stress is a novel application in wood material science. The method offers the potential for the simultaneous observation of structural changes and quantified deformations during in situ micro-mechanical experiments. Moreover, the high spatial resolution allows studying the influence of anatomical features on the fracture behaviour of wood. Possible applications of this method range from bio-mechanical observations in fresh plant tissue to fracture mechanics aspects in structural timber.

Monitoring angiogenesis in human melanoma xenograft model using contrast-enhanced ultrasound imaging.

The potential for noninvasive monitoring and quantification of tumor angiogenesis with contrast-enhanced ultrasound imaging has been investigated in a murine cancer model. Seventy athymic nude mice were implanted with the human melanoma cell line DB-1 but only 30 of these were available for the final study. The 30 mice were divided into three groups (10 mice/group), which were studied with contrast-enhanced ultrasound imaging 4, 5 or 6 weeks post-implantation. Power Doppler and pulse inversion harmonic imaging (PIHI) were performed (in real time and intermittently) with a Sonoline Elegra scanner (Siemens Medical Solutions, Issaquah, WA) following injection of Optison (dose: 0.4-0.6 ml/kg; GE Healthcare, Princeton, NJ). Ultrasound results were compared to immunohistochemical stains for endothelial cells (CD31), vascular endothelial growth factor (VEGF) and cyclooxygenase-2 (COX-2). Linear regression analysis indicated statistically significant correlations between the percent area stained with VEGF and ultrasound measures of tumor neovascularity obtained with all three techniques (p < 0.01). Contrast-enhanced ultrasound imaging of tumor neovascularity appears to provide a noninvasive marker of angiogenesis corresponding to the expression of VEGF in the DB-1 model and may become a useful tool for monitoring clinical anti-angiogenic therapies.

Application of the compound probability density function for characterization of breast masses in ultrasound B scans.

The compound probability density function (pdf) is investigated for the ability of its parameters to classify masses in ultrasonic B scan breast images. Results of 198 images (29 malignant and 70 benign cases and two images per case) are reported and compared to the classification performance reported by us earlier in this journal. A new parameter, the speckle factor, calculated from the parameters of the compound pdf was explored to separate benign and malignant masses. The receiver operating characteristic curve for the parameter resulted in an A(z) value of 0.852. This parameter was combined with one of the parameters from our previous work, namely the ratio of the K distribution parameter at the site and away from the site. This combined parameter resulted in an A(z) value of 0.955. In conclusion, the parameters of the K distribution and the compound pdf may be useful in the classification of breast masses. These parameters can be calculated in an automated fashion. It should be possible to combine the results of the ultrasonic image analysis with those of traditional mammography, thereby increasing the accuracy of breast cancer diagnosis.

Ultrasonic biomedical technology; marketing versus clinical reality.

Clinical ultrasound imaging is the most frequently used imaging modality in the world accounting for almost 25% of all imaging studies performed. Hence, manufacturers of commercial ultrasound equipment are committing significant engineering resources to advance the technology behind the modality and to improve the diagnostic capabilities of ultrasound imaging. Consequently, new imaging technologies are constantly being introduced to the market under a host of different trademarked names. Hence, this paper will review many of the recent advances in ultrasound imaging and try to differentiate between developments aimed at marketing and those providing real clinical improvements. In particular, we will describe the technologies behind concepts such as coded ultrasound imaging, real time spatial compounding, tissue harmonic imaging, extended field of view imaging as well as 3D and 4D (i.e., real time 3D) imaging. Some of the latest advances in blood flow imaging e.g., B-flow, advanced dynamic flow (ADF) and automatic optimization methods will also be described.

Assessment of angiogenesis: implications for ultrasound imaging.

In this paper, the fundamentals of tumor angiogenesis and the implications for ultrasound imaging will be described. Twenty-eight athymic nude mice were implanted with the human melanoma cell lines DB-1 or MW-9 (14 mice/group). Ultrasound contrast agents were injected in the tail veins. Power Doppler and pulse inversion harmonic imaging (PI-HI) was performed (in real time and intermittently). Ultrasound results were compared to immunohistochemical stains for endothelial cells (CD31), vascular endothelial growth factor (VEGF), and cyclooxygenase-2 (COX-2). Linear regression analysis indicated statistically significant correlations between percent area stained with COX-2 and with VEGF relative to power Doppler (p<0.05) and intermittent PI-HI (p<0.05) measures of tumor neovascularity in the MW-9 and the DB-1 mice, respectively. Preliminary results from a human trial of the anti-angiogenic drug Angiostatin (Entremed, Rockville, MD) showed tumor volumes increased in two patients, while the vascularity remained virtually unchanged. Conversely, in three patients with diminished tumor volumes vascularity increased by 38%. In conclusion, contrast enhanced ultrasound imaging of tumor neovascularity may provide noninvasive markers of angiogenesis and may become a useful tool for monitoring anti-angiogenic therapies in vivo.

Acoustic microscopy system: design and preliminary results.

An acoustic microscopy system was designed to perform 2D imaging in the C-plane with a single-element transducer. The ultrasound transducer was fabricated by polishing bulk lithium niobate (LiNbO(3)) to the required thickness (approximately 60 or 45 micro) for the desired operating frequency (55 or 75 MHz). The polished LiNbO(3) was attached to acoustic backing and matching layers. Finally, an epoxy lens was applied and the transducer mounted in a housing. The transducer was mounted in a 3D motorized positioning stage and operated by a high-frequency pulser/receiver. Received echoes were sampled with a 2 GHz ADC card and displayed on a PC using software developed in the Matlab environment. Transducer frequency and bandwidth were measured off a steel plate positioned at the focal length. A penny was scanned initially to confirm expected performance before acquiring data from liver (n=3) and spleen (n=3) specimens. For the first probe, the peak frequency was 54.05 MHz with a -6 dB bandwidth of 6.76 MHz. The axial and lateral resolutions were estimated to be 114 and 188 microm, respectively. For the second probe, the peak frequency was measured to 82 MHz with a -6 dB bandwidth of approximately 23 MHz. The axial and lateral resolutions were estimated to be around 33 and 81 microm, respectively. C-scans of the penny clearly showed detailed structures on front and back, while the capsule and the trabecular structures of the splenic tissues could easily be separated in different layers. In conclusion, an acoustic microscopy system operating at 55-75 MHz has been constructed and the feasibility of obtaining high-resolution images of tissue specimens demonstrated.

Classification of breast masses in ultrasonic B-mode images using a compounding technique in the Nakagami distribution domain.

Classification of masses in ultrasonic B-mode images of the breast tissue using "normalized" parameters of the Nakagami distribution was recently investigated. The technique, however, did not yield performances that were comparable to those of an experienced radiologist, and utilized only a single image for tissue characterization. Because radiologists commonly use two to four images of a mass for characterization, a similar procedure is developed here. A simple summation of the normalized Nakagami parameters from two different images of a mass is utilized for classification as benign or malignant. The performance of the normalized Nakagami parameters before and after the summation has been carried out through a receiver operating characteristic (ROC) study. The bootstrap procedure has been utilized to compute the mean and SD of the ROC area, A(z), obtained for each parameter. It has been observed that combining normalized Nakagami parameters from two images of the mass may help to improve classification performance over that from utilizing the parameters of just a single image. The performance of this automated parameter-based approach appears to match that of a trained radiologist.

Computer aided classification of masses in ultrasonic mammography.

Frequency compounding was recently investigated for computer aided classification of masses in ultrasonic B-mode images as benign or malignant. The classification was performed using the normalized parameters of the Nakagami distribution at a single region of interest at the site of the mass. A combination of normalized Nakagami parameters from two different images of a mass was undertaken to improve the performance of classification. Receiver operating characteristic (ROC) analysis showed that such an approach resulted in an area of 0.83 under the ROC curve. The aim of the work described in this paper is to see whether a feature describing the characteristic of the boundary can be extracted and combined with the Nakagami parameter to further improve the performance of classification. The combination of the features has been performed using a weighted summation. Results indicate a 10% improvement in specificity at a sensitivity of 96% after combining the information at the site and at the boundary. Moreover, the technique requires minimal clinical intervention and has a performance that reaches that of the trained radiologist. It is hence suggested that this technique may be utilized in practice to characterize breast masses.

Contrast enhanced vascular three-dimensional ultrasound imaging.

In other imaging modalities three-dimensional (3D) data displays are well established; not so in ultrasound. Due to the real-time requirements of ultrasound the time available to compute 3D displays is limited, particularly when flow data is acquired with Doppler techniques. Consequently, it is only recently that improvements in computer processing power have resulted in useful vascular 3D ultrasound scans. Many manufacturers have now implemented free-hand 3D power Doppler capabilities on their scanners. However, to obtain flow signals from smaller vessels associated e.g., with tumor neovascularity, may very well require the introduction of a microbubble based ultrasound contrast agent into the blood stream. Given the up to 30 dB enhancement of Doppler signals produced by the contrast microbubbles quite spectacular vascular 3D images are feasible. Moreover, new contrast imaging techniques, such as harmonic imaging, have now permitted 3D vascular information to be acquired and displayed in grayscale with the associated improvement in resolution. In this paper we will review different aspects of contrast enhanced vascular 3D ultrasound imaging including implementation, contrast specific techniques and in vivo imaging.

Classification of ultrasonic B mode images of the breast using frequency diversity and Nakagami statistics.

The parameters of the Nakagami distribution have been utilized in the past to classify lesions in breast tissue as benign or malignant. To avoid the effect of operatorgain settings on the parameters of the Nakagami distribution, normalized parameters were utilized for the classification. The normalized parameter was defined as the ratio of the parameter at the site of the lesion to its average value over several regions away from the site. This technique, however, was very time consuming. In this paper, the application of frequency diversity and compounding is explored to achieve this normalization. Lesions are classified using these normalized parameters at the site. A receiver operating characteristic (ROC) analysis of the parameters of the Nakagami distribution has been conducted before and after compounding on a data set of 60 benign and 65 malignant lesions. The ROC results indicate that this technique can reasonably classify lesions in breast tissue as benign or malignant.

Tissue classification with generalized spectrum parameters.

This paper presents performance comparisons between breast tumor classifiers based on parameters from a conventional texture analysis (CTA) and the generalized spectrum (GS). The computations of GS-based parameters from radiofrequency (RF) ultrasonic scans and their relationship to underlying scatterer properties are described. Clinical experiments demonstrate classifier performances using 22 benign and 24 malignant breast mass regions taken from 40 patients. Linear classifiers based on parameters from the front edge, back edge and interior tumor regions are examined. Results show significantly better performances for GS-based classifiers, with improvements in empirical receiver operating characteristic (ROC) areas of greater than 10%. The ROC curves show GS-based classifiers achieving a 90% sensitivity level at 50% specificity when applied to the back-edge tumor regions, an 80% sensitivity level at 65% specificity when applied to the front-edge tumor regions, and a 100% sensitivity level at 45% specificity when applied to the interior tumor regions.