KEYWORDS: Tumors, Tissues, Control systems, Signal attenuation, Prostate, Prostate cancer, Ultrasonography, Tumor growth modeling, Transducers, In vivo imaging
Optoacoustic (OA) imaging was employed to distinguish normal from neoplastic tissues in a transgenic murine model of prostate cancer. OA images of five tumor-bearing mice and five age-matched controls across a 14 mm×14 mm region of interest (ROI) on the lower abdomen were acquired using a reverse-mode OA imaging system (Seno Medical Instruments Inc., San Antonio, Texas). Neoplastic prostate tissue was identified based on the OA signal amplitude in combination with spectral analysis of the OA radio frequency (RF) data. Integration of the signal amplitude images was performed to construct two-dimensional images of the ROI. The prostate tumors generated higher amplitude signals than those of the surrounding tissues, with contrast ratios ranging from 31 to 36 dB. The RF spectrum analysis showed significant differences between the tumor and the control mice. The midband fit was higher by 5 dB (62%), the intercept higher by 4 dB (57%) and the spectral slope higher by 0.4 dB/MHz (50%) for neoplastic prostate tissue compared to normal tissues in the control mice. The results demonstrate that OA offers high contrast imaging of prostate cancer in vivo.
The canine prostate is a close match for the human prostate and is used in research of prostate cancers. Determining accurately optical absorption and scattering properties of the gland in a wide spectral range (preferably in a minimally invasive way), linking optical properties to concentrations of major endogenous chromophores, and detecting the presence of localized optical inhomogeneities like inclusions of gold nanoparticles for therapeutic and diagnostic purposes, are among the major challenges for researchers. The goal of the article is to demonstrate a feasibility of the multifunctional radiance spectroscopy platform in providing the required information. For ex vivo canine prostate, extraction of the effective attenuation and diffusion coefficients using relative cw radiance measurements was demonstrated in the 650- to 900-nm range. The derived absorption coefficient was decomposed to contributions from 9.0 μM HbO2, 29.6 μM Hb, and 0.47 fractional volume of H2O. Detection of a localized inclusion containing ∼1.5·1010 gold nanorods (0.8 μg Au) at 10 mm distance from the urethra was achieved with the detector in the urethra and the light source in a virtual rectum position. The platform offers the framework for a systematic study of various chromophores in the prostate that can be used as comprehensive diagnostic markers.
Gold nanoparticles can be used as molecular contrast agents binding specifically to cancer sites and thus delineating tumor regions. Imaging gold nanoparticles deeply embedded in tissues with optical techniques possesses significant challenges due to multiple scattering of optical photons that blur the obtained images. Both diagnostic and therapeutic applications can benefit from a minimally invasive technique that can identify, localize, and quantify the payloads of gold nanoparticles deeply embedded in biological tissues. An optical radiance technique is applied to map localized inclusions of gold nanorods in 650- to 900-nm spectral range in a porcine phantom that mimics prostate geometry. Optical radiance defines a variation in the angular density of photons impinging on a selected point in the tissue from various directions. The inclusions are formed by immersing a capillary filled with gold nanorods in the phantom at increasing distances from the detecting fiber. The technique allows the isolation of the spectroscopic signatures of the inclusions from the background and identification of inclusion locations in the angular domain. Detection of ∼4×10 10 gold nanoparticles or 0.04 mg Au/mL (detector–inclusion separation 10 mm, source–detector separation 15 mm) in the porcine tissue is demonstrated. The encouraging results indicate a promising potential of radiance spectroscopy in early prostate cancer diagnostics with gold nanoparticles.
We have applied an optical radiance technique to map localized inclusions of gold nanoparticles in a porcine phantom. Our goal was to show that combined spectroscopic and angular snapshots of phantoms allow the obtaining of information that is relevant for prostate cancer diagnostics. A combination of the radiance spectroscopy and white light spectroscopy was used to measure angular resolved light distribution in 600-900 nm spectral range inside the porcine phantom that mimics prostate geometry. Optical radiance defines a variation in the angular density of photons impinging on a selected point in the tissue from various directions. To obtain radiance data, a specially constructed optical probe with a well-defined angular detection window must be rotated along its axis. Characteristic spectro-angular snapshots of the phantom alone and with the localized inclusion of gold nanoparticles were obtained. The inclusions were formed by immersing a capillary filled with gold nanoparticles into selected locations in the phantom. For phantoms with gold inclusions, this approach allows the isolation of the spectroscopic signatures of the inclusions from the background and identification of inclusion locations in the angular domain. Detection of ~1010 gold nanoparticles (detector-inclusion separation 10 mm, source-detector separation 15 mm) in the porcine tissue was demonstrated. These encouraging results indicate a promising potential of radiance spectroscopy in early prostate cancer diagnostics with gold nanoparticles.
We present a new approach to the analysis of radiance in turbid media. The approach combines data from spectral, angular and spatial domains in a form of spectro-angular maps. Mapping provides a unique way to visualize details of light distribution in turbid media and allows tracking changes with distance. Information content of experimental spectro-angular maps is verified by a direct comparison with simulated data when an analytical solution of the radiative transfer equation is used. The findings deepen our understanding of the light distribution in a homogenous turbid medium and provide a first step toward applying the spectro-angular mapping as a diagnostic tool for tissue characterization.
We have applied an interstitial radiance-based technique based on a spectro-angular mapping approach to the
identification and angular localization of 250-nm and 5-nm Au nanoparticle-based inclusions and non-scattering (water
only) inclusions in the Intralipid-1% liquid phantom. A combination of the point radiance spectroscopy and white light
spectroscopy was used to measure angular resolved light distribution in 450-900 nm spectral range in Intralipid-1% with
and without localized inclusions. Characteristic spectro-angular snapshots of the liquid phantom alone and with the
localized inclusions were obtained. For liquid phantoms without inclusions, the snapshots demonstrate wavelength
dependent light distribution inside the turbid medium. For liquid phantoms with gold inclusions, the approach allows to
isolate the spectroscopic signatures of the inclusions from the background, identify locations of the inclusions in the
angular domain and show how a presence of water in the inclusion affects spectral identification and angular localization
of the target. For liquid phantoms with water-based inclusions, an ability of the inclusion to enhance photon density
above reference values and angular dependent signatures were demonstrated. The technique is seen as a potential tool in
prostate treatment and diagnostics with gold nanoparticles.
Laser thermal therapy involves heating tissue using light to temperatures between 55 °C and 95 °C for several minutes
resulting in coagulation and cell death. This treatment method has been under investigation for use as a minimally
invasive method for treating solid tumors and cancer cells. Heating tissues results in highly variable outcomes and
challenges; for example, ensuring complete coagulation of the target tissue while avoiding damage to surrounding
healthy tissues. Overcoming such challenges requires precise and real-time monitoring. Optoacoustic imaging has been
proposed as a real-time, noninvasive method for monitoring laser thermal. Ex-vivo porcine tenderloin samples were
heated using a 1000 μm core optical fiber coupled to an 810 nm diode laser at a constant power of 7 W for 10 minutes.
Lesions (6-7 mm diameter) were scanned using a prototype
reverse-mode optoacoustic system consisting of a pulsed
laser which operates at 1064 nm coupled to a bifurcated fibre bundle, and an 8 element annular array wideband
ultrasound transducer with a central frequency ~5 MHz. Scanning was done across native and coagulated tissue with an
energy of 6.5 mJ at a 1064 nm wavelength. Three lesions of similar size, shape and coagulation state were chosen for
analysis. Thermal coagulation effects were analyzed using optoacoustic signal amplitude and spectral analysis of the
optoacoustic RF data. Results show that the signal amplitude and the intercept and midband fit of the power spectrum
exhibit interesting differences between native and coagulated tissue states.
Interstitial fiber-optic-based approaches used in both diagnostic and therapeutic applications rely on localized light-tissue interactions. We present an optical technique to identify spectrally and spatially specific exogenous chromophores in highly scattering turbid media. Point radiance spectroscopy is based on directional light collection at a single point with a side-firing fiber that can be rotated up to 360 deg. A side firing fiber accepts light within a well-defined, solid angle, thus potentially providing an improved spatial resolution. Measurements were performed using an 800-μm diameter isotropic spherical diffuser coupled to a halogen light source and a 600 μm, ∼43 deg cleaved fiber (i.e., radiance detector). The background liquid-based scattering phantom was fabricated using 1% Intralipid. Light was collected with 1 deg increments through 360 deg-segment. Gold nanoparticles , placed into a 3.5-mm diameter capillary tube were used as localized scatterers and absorbers introduced into the liquid phantom both on- and off-axis between source and detector. The localized optical inhomogeneity was detectable as an angular-resolved variation in the radiance polar plots. This technique is being investigated as a potential noninvasive optical modality for prostate cancer monitoring.
The interferometric measurement of laser induced thermoelastic expansion of tissue samples can be used to estimate
their optical, thermal and mechanical properties. This method was used to assess the Gruneisen coefficient and optical
attenuation depth for native and coagulated ex-vivo bovine liver and porcine kidney samples. The results demonstrate
decreases of 54% and 60% in the optical attenuation depth in bovine liver and porcine kidney after coagulation,
respectively. The Gruneisen coefficient of native porcine kidney was determined to be 58% smaller (p < 0.05) that native
bovine liver. The measured Gruneisen coefficients for native and coagulated ex-vivo porcine kidney were 0.07 ± 0.03
and 0.105 ± 0.02, respectively, whereas the Gruneisen coefficients for native and coagulated liver were 0.126 ± 0.036
and 0.127 ± 0.04, respectively. Our measurements indicate significant inter sample variability due likely to inherent
variations in tissue optical absorption and surface preparation.
Interstitial fiber-optic based strategies for therapy monitoring and assessment rely on detecting treatment-induced
changes in the light distribution in biological tissues. We present an optical technique to identify spectrally and spatially
specific tissue chromophores in highly scattering turbid media. Typical optical sensors measure non-directional light
intensity (i.e. fluence) and require fiber translation (i.e. 3-5 positions), which is difficult to implement clinically. Point
radiance spectroscopy is based on directional light collection (i.e. radiance) at a single point with a side-firing fiber that
can be rotated up to 360°. A side firing fiber accepts light within a well-defined solid angle thus potentially providing an
improved spatial resolution. Experimental measurements were performed using an 800-μm diameter isotropic spherical
diffuser coupled to a halogen light source and a 600 μm, ~43° cleaved fiber (i.e. radiance detector). The background
liquid-based scattering phantom was fabricated using 1% Intralipid (i.e. scattering medium). Light was collected at 1-5° increments through 360°-segment. Gold nanoparticles, placed into a 3.5 mm diameter capillary tube were used as
localized scatterers and absorbers introduced into the liquid phantom both on- and off-axis between source and detector.
The localized optical inhomogeneity was detectable as an angular-resolved variation in the radiance polar plots. This
technique is being investigated as a non-invasive optical modality for prostate cancer monitoring.
Several studies have explored the potential of optoacoustic imaging for monitoring thermal therapies, yet the origin of the contrast in the images is not well understood. A technique is required to measure the changes in the optical and thermomechanical properties of tissues upon coagulation to better understand this contrast. An interferometric method is presented for measuring simultaneously the optical and thermomechanical properties of native and coagulated ex-vivo bovine tissue samples based on analysis of the surface displacement of irradiated samples. Surface displacements are measured after irradiation by short laser pulses at 750 nm. A 51% decrease in the optical attenuation depth is observed for coagulated liver samples compared to native samples. No significant differences in the Grüneisen coefficient are measured in the native and coagulated tissue samples. A mean value of 0.12 for the Grüneisen coefficient is measured for both native and coagulated liver tissues. The displacement profiles exhibit consistent differences between the two tissue types. To assess the changes in the sample mechanical properties, the experimental data also are compared to numerical solutions of the equation for thermoelastic deformation. The results demonstrate that differences in the tissue expansion dynamics arise from higher values of elastic modulus for coagulated liver samples compared to native ones.
Raman spectroscopy is used to study the effects of heating on specific molecular bonds present in albumen-based coagulation phantoms and ex-vivo tissues. Thermal coagulation is induced by submerging albumen-based phantoms in a 75°C water bath to achieve target temperatures of 45, 55, 65, and 75°C. Laser photocoagulation is performed on ex-vivo bovine muscle samples, yielding induced temperatures between 46 and 90°C, as reported by implanted microthermocouples. All phantoms and tissue samples are cooled to room temperature, and Raman spectra are acquired at the microthermocouple locations. Shifts in major Raman bands are observed with laser heating in bovine muscle, specifically from the amide-1 -helix group (~1655 cm−1), the CH2/CH3 group (~1446 cm−1), the C-H stretch group (~1312 cm−1), and the CN stretch group (~1121cm−1). Raman bands at 1334 cm−1 (tryptophan), 1317 cm−1 [(C-H)], and 1655 cm−1 (amide-1 -helix) also show a decrease in intensity following heating. The results suggest that Raman band locations and relative intensities are affected by thermal denaturation of proteins, and hence, may be a useful tool for monitoring the onset and progression of coagulation during thermal therapies.
Prostate cancer is currently the most common cancer among Canadian men. Due to an increase in public awareness and
screening, prostate cancer is being detected at earlier stages and in much younger men. This is raising the need for better
treatment monitoring approaches. Optoacoustic imaging is a new technique that involves exposing tissues to pulsed light
and detecting the acoustic waves generated by the tissue. Optoacoustic images of a tumour bearing mouse and an agematched
control were acquired for a 775 nm illumination using a reverse-mode imaging system. A murine model of
prostate cancer, TRAMP (transgenetic adenocarcinoma of mouse prostate), was investigated. The results show an
increase in optoacoustic signal generated by the tumour compared to that generated by the surrounding tissues with a
contrast ratio of 3.5. The dimensions of the tumour in the optoacoustic image agreed with the true tumour dimensions to
within 0.5 mm. In this study we show that there are detectable changes in optoacoustic signal strength that arise from
the presence of a tumour in the prostate, which demonstrates the potential of optoacoustic imaging for the monitoring of
prostate cancer therapy.
Dynamics of the thermoelastic expansion of native and coagulated
ex-vivo bovine liver tissues after their irradiation
by short laser pulses were studied. The differences in optical and thermo-mechanical properties of the native and
coagulated samples such as their Gruneisen coefficient and optical attenuation depth were quantitatively determined. It
was found that for coagulated ex-vivo bovine liver samples, the optical attenuation depth decrease by an average of 47%.
Also significant differences were observed in the dynamics of thermoelastic expansion of the tissue surface. These
differences can be potentially linked to differences in
thermo-mechanical properties between native and coagulated
samples. The changes in these properties may be
Optoacoustic imaging is a relatively novel biomedical imaging modality that relies on the absorption of light to create
pressure transients that can be detected ultrasonically. In most scientific communications, the source of tissue contrast
has been described as primarily optical. However, the thermomecahnical properties of tissue, as expressed through the
Gruneisen coefficient, also affect the optoacoustic signal. To investigate the effect of thermomechanical tissue properties
short pulses (~ 6.5 ns) from an optical parametric oscillator at 750 nm were used to irradiate coagulated and
uncoagulated tissue-mimicking albumen phantoms, to emulate normal tissue and tissue that has been heated. The
phantoms respond to the laser-induced stress by thermoelastic expansion. This thermomechanical behavior of the
samples was assessed using an interferometric system capable of measuring transient displacements with a temporal
resolution of less than 10 ns and a spatial resolution of < 10 nm. The experimental measurement allowed determination
of the Gruneisen coefficient which is an important thermo-mechanical sample property that can affect generation of
optoacoustic signals. An increase in the value of Gruneisen coefficient of 65% was measured when phantoms were
coagulated compared to uncoagulated phantoms, consistent with the stiffening of the tissue mimicking material. This
suggests that for thermal therapy the changes in the Gruneisen coefficient are also an important source of optoacoustic
contrast.
Minimally invasive thermal therapy is being investigated as an alternative cancer treatment. It involves heating
tissues to greater than 55°C over a period of a few minutes, which results in tissue coagulation. Optoacoustic (OA)
imaging is a new imaging technique that involves exposing tissues to pulsed light and detecting the acoustic waves
that are generated. In this study, adult bovine liver tissue samples were heated using continuous wave laser energy
for various times, then scanned using an optoacoustic imaging system. Large optoacoustic signal variability was
observed in the native tissue prior to heating. OA signal amplitude increased with maximum tissue temperature
achieved, characterized by a correlation coefficient of 0.63. In this study we show that there are detectable changes
in optoacoustic signal strength that arise from tissue coagulation, which demonstrates the potential of optoacoustic
technology for the monitoring of thermal therapy delivery.
Photoacoustic tomography provides good optical contrast with high spatial resolution making it an attractive tool for
noninvasive imaging. While the mechanical parameters of tissue affect the photoacoustic signal, the differences in
optical absorption mainly determines the contrast between different media. In this work we investigate how the variation
in optical and mechanical properties during laser-induced coagulation can be detected by changes in the amplitude and
temporal characteristics of photoacoustic signals. Photoacoustic pressure profiles are investigated for tissue equivalent
albumen phantoms exposed to varying thermal doses, simulating thermal coagulation. Illumination is performed using
an optical parametric oscillator (OPO) fed by a Q-switched Nd:YAG pulsed laser to illuminate at multiple wavelengths.
The results of the study demonstrate that photoacoustic signals are sensitive to changes in delivered thermal dose and,
hence, photoacoustic imaging has potential as a non-invasive monitoring tool for thermal therapy.
Interstitial quantification of the optical properties of tissue is important in biomedicine for both treatment planning of minimally invasive laser therapies and optical spectroscopic characterization of tissues, for example, prostate cancer. In a previous study, we analyzed a method first demonstrated by Dickey et al., [Phys. Med. Biol. 46, 2359 (2001)] to utilize relative interstitial steady-state radiance measurements for recovering the optical properties of turbid media. The uniqueness of point radiance measurements were demonstrated in a forward sense, and strategies were suggested for improving performance under noisy experimental conditions. In this work, we test our previous conclusions by fitting the P3 approximation for radiance to Monte Carlo predictions and experimental data in tissue-simulating phantoms. Fits are performed at: 1. a single sensor position (0.5 or 1 cm), 2. two sensor positions (0.5 and 1 cm), and 3. a single sensor position (0.5 or 1 cm) with input knowledge of the sample's effective attenuation coefficient. The results demonstrate that single sensor radiance measurements can be used to retrieve optical properties to within ~20%, provided the transport albedo is greater than ~0.9. Furthermore, compared to the single sensor fits, employing radiance data at two sensor positions did not significantly improve the accuracy of recovered optical properties. However, with knowledge of the effective attenuation coefficient of the medium, optical properties can be retrieved experimentally to within ~10% for an albedo greater or equal to 0.5.
Optoacoustic Imaging is based on the thermal expansion of tissue caused by a temperature rise due to absorption of short laser pulses. At constant laser fluence, optoacoustic image contrast is proportional to differences in optical absorption and the thermoacoustic efficiency, expressed by the Grueuneisen parameter, Γ. Γ is proportional to the thermal expansion coefficient, the sound velocity squared and the inverse heat capacity at constant pressure. In thermal therapies, these parameters may be modified in the treated area. In this work experiments were performed to examine the influence of these parameters on image contrast. A Laser Optoacoustic Imaging System (LOIS, Fairway Medical Technologies, Houston, Texas) was used to image tissue phantoms comprised of cylindrical Polyvinyl Chloride Plastisol (PVCP) optical absorbing targets imbedded in either gelatin or PVCP as the background medium. Varying concentrations of Black Plastic Color (BPC) and titanium dioxide (TiO2) were added to targets and background to yield desired tissue relevant optical absorption and effective scattering coefficients, respectively. In thermal therapy experiments, ex-vivo bovine liver was heated with laser fibres (805nm laser at 5 W for 600s) to create regions of tissue coagulation. Lesions formed in the liver tissue were visible using the LOIS system with reasonable correspondence to the actual region of tissue coagulation. In the phantom experiments, contrast could be seen with low optical absorbing targets (μa of 0.50cm-1 down to 0.13cm-1) embedded in a gelatin background (see manuscript for formula). Therefore, the data suggest that small objects (< 5mm) with low absorption coefficients (in the range < 1cm-1) can be imaged using LOIS. PVCP-targets in gelatin were visible, even with the same optical properties as the gelatin, but different Γ. The enhanced contrast may also be caused by differences in the mechanical properties between the target and the surrounding medium. PVCP-targets imbedded in PVCP produced poorer image contrast than PVCP-targets in gelatin with comparable optical properties. The preliminary investigation in tissue equivalent phantoms indicates that in addition to tissue optical properties, differences in mechanical properties between heated and unheated tissues may be responsible for image contrast. Furthermore, thermal lesions in liver tissue, ex-vivo, can be visualized using an optoacoustic system.
Laser Optoacoustic Imaging System (LOIS) combines high tissue contrast based on the optical properties of tissue and high spatial resolution based on ultrawide-band ultrasonic detection. Patients undergoing thermal or photodynamic therapy of prostate cancer may benefit from capability of LOIS to detect and monitor treatment-induced changes in tissue optical properties and blood flow. The performance of a prototype LOIS was evaluated via 2D optoacoustic images of dye-colored objects of various shapes, small tubes with blood simulating veins and arteries, and thermally coagulated portions of chicken breasts imbedded tissue-mimicking gelatin phantoms. The optoacoustic image contrast was proportional to the ratio of the absorption coefficient between the embedded objects and the surrounding gel. The contrast of the venous blood relative to the background exceeded 250%, and the contrast of the thermally coagulated portions of flesh relative to the untreated tissue ranged between -100% to +200%, dependent on the optical wavelength. We used a 32-element optoacoustic transducer array and a novel design of low-noise preamplifiers and wide-band amplifiers to perform these studies. The system was optimized for imaging at a depth of ~50 mm. The system spatial resolution was better than 1-mm. The advantages and limitations of various signal-processing methods were investigated. LOIS demonstrates clinical potential for non- or minimally-invasive monitoring of treatment-induced tissue changes.
Frequency-domain correlation and spectral analysis photothermoacoustic (FD-PTA) imaging is a promising new technique, which is being developed to detect tumor masses in turbid biological tissue. Unlike conventional biomedical photoacoustics which uses time-of-flight acoustic information induced by a pulsed laser to indicate the tumor size and location, in this research, a new FD-PTA instrument featuring frequency sweep (chirp) and heterodyne modulation and lock-in detection of a continuous-wave laser source at 1064 nm wavelength is constructed and tested for its depth profilometric capabilities in turbid media imaging. Owing to the linear relationship between the depth of acoustic signal generation and the delay time of signal arrival to the transducer, information specific to a particular depth can be associated with a particular frequency in the chirp signal. Scanning laser-fluence modulation frequencies with a linear frequency sweep method preserves the depth-to-delay time linearity and recovers FD-PTA signals from a range of depths. A report on two-dimensional spatial scans, performed on tissue mimicking control phantoms with various optical, acoustical and geometrical properties will be presented. Combining with the depth information carried by the back-propagated chirp signal at each scanning position, one could rapidly generate sub-surface three-dimensional images of the scanning area, a combination of tasks that is difficult or impossible by use of pulsed photoacoustic detection. It is concluded that frequency domain photothermoacoustics using a linear frequency sweep method and heterodyne lock-in detection has the potential to be a reliable tool for biomedical depth-profilometric imaging.
Laser thermal therapy (LTT) is a minimally invasive surgical technique used to destroy solid tumors while minimizing damage to adjacent normal tissues. Optical energy, delivered through fibers implanted into the target volume, raises tissue temperatures above 60 degree(s)C resulting in coagulative necrosis (thermal damage). Thermal damage volumes, however, can be irregular and unpredictable, resulting from dynamic changes in the tissue properties during treatment. A closed-loop feedback fuzzy logic controller for LTT was developed with the tissue treated as a black-box system. Preliminary testing was conducted for simulated LTT with a single spherically emitting source fiber at the center of 5 mm and 10 mm diameter target tissues. Dynamic changes in blood perfusion and tissue optical properties due to heating were incorporated into the LTT simulator. Input laser power was modulated to control the temperature field in an attempt to reach target temperatures at the source (90 degree(s)C to avoid tissue charring) and at the target boundary (55 degree(s)C). In all simulations, thermal damage based on Arrhenius formulation ((Omega) equals 1) was reached at the target boundary. The controller also responded efficiently to unexpected, rapid temperature changes.
The construction of a software platform for planning of photodynamic therapy (PDT) treatments is described. Its use is illustrated by the planning of a treatment of prostate cancer using multiple interstitial optical fiber sources. The effects of changing tissue optical properties, of photosensitizer photobleaching and of altering the power distribution between the sources are demonstrated. Validation tests of the PDT dose calculation algorithms and the software implementation of these are included. The possibility of compensating for misplaced sources is shown and confirmed experimentally in a tissue-simulating phantom. The future development of this platform and its integration into a complete solution planning-delivery-control platform is discussed.
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