Samuel Pichardo

Multi-frequency characterization of the speed of sound and attenuation coefficient for longitudinal transmission of freshly excised human skulls

For medical applications of ultrasound inside the brain, it is necessary to understand the relationship between the apparent density of skull bone and its corresponding speed of sound and attenuation coefficient. Although there have been previous studies exploring this phenomenon, there is still a need to extend the measurements to cover more of the clinically relevant frequency range. The results of measurements of the longitudinal speed of sound and attenuation coefficient are presented for specimens of human calvaria. The study was performed for the frequencies of 0.27, 0.836, 1.402, 1.965 and 2.525 MHz. Specimens were obtained from fresh cadavers through a protocol with the Division of Anatomy of the University of Toronto. The protocol was approved by the Research Ethics Board of Sunnybrook Health Sciences Centre. The specimens were mounted in polycarbonate supports that were marked for stereoscopic positioning. Computer tomography (CT) scans of the skulls mounted on their supports were performed, and a three-dimensional skull surface was reconstructed. This surface was used to guide a positioning system to ensure the normal sound incidence of an acoustic signal. This signal was produced by a focused device with a diameter of 5 cm and a focal length of 10 cm. Measurements of delay in time of flight were carried out using a needle hydrophone. Measurements of effective transmitted energy were carried out using a radiation force method with a 10 µg resolution scale. Preliminary functions of speed of sound and attenuation coefficient, both of which are related to apparent density, were established using a multi-layer propagation model that takes into account speed of sound, density and thickness of the layer. An optimization process was executed from a large set of random functions and the best functions were chosen for those ones that closest reproduced the experimental observations. The final functions were obtained after a second pass of the optimization process was executed, but this time using a finite-difference time-difference solution of the Westervelt equation, which is more precise than the multi-layer model but much more time consuming for computation. For six of seven specimens, measurements were carried out on five locations on the calvaria, and for the other specimen three measurements were made. In total, measurements were carried out on 33 locations. Results indicated the presence of dispersion effects and that these effects are different according to the type of bone in the skull (cortical and trabecular). Additionally, both the speed of sound and attenuation showed dependence on the skull density that varied with the frequency. Using the optimal functions and the information of density from the CT scans, the average values (±s.d.) of the speed of sound for cortical bone were estimated to be 2384(± 130), 2471(± 90), 2504(± 120), 2327(± 90) and 2053(± 40) m s(-1) for the frequencies of 270, 836, 1402, 1965 and 2526 kHz, respectively. For trabecular bone, and in the same order of frequency values, the speeds of sound were 2140(± 130), 2300(± 100), 2219(± 200), 2133(± 130) and 1937(± 40) m s(-1), respectively. The average values of the attenuation coefficient for cortical bone were 33(± 9), 240(± 9) and 307(± 30) Np m(-1) for the frequencies of 270, 836, and 1402, respectively. For trabecular bone, and in the same order of frequency values, the average values of the attenuation coefficient were 34(± 13), 216(± 16) and 375(± 30) Np m(-1), respectively. For frequencies of 1.965 and 2.525 MHz, no measurable radiation force was detected with the setup used.

High intensity focused ultrasound technology, its scope and applications in therapy and drug delivery.

Ultrasonography is a safe, inexpensive and wide-spread diagnostic tool capable of producing real-time non-invasive images without significant biological effects. However, the propagation of higher energy, intensity and frequency ultrasound waves through living tissues can induce thermal, mechanical and chemical effects useful for a variety of therapeutic applications. With the recent development of clinically approved High Intensity Focused Ultrasound (HIFU) systems, therapeutic ultrasound is now a medical reality. Indeed, HIFU has been used for the thermal ablation of pathological lesions; localized, minimally invasive ultrasound-mediated drug delivery through the transient formation of pores on cell membranes; the temporary disruption of skin and the blood brain barrier; the ultrasound induced break-down of blood clots; and the targeted release of drugs using ultrasound and temperature sensitive drug carriers. This review seeks to engage the pharmaceutical research community by providing an overview on the biological effects of ultrasound as well as highlighting important therapeutic applications, current deficiencies and future directions.

Sonoporation delivery of monoclonal antibodies against human papillomavirus 16 E6 restores p53 expression in transformed cervical keratinocytes.

High-risk types of human papillomavirus (HPV), such as HPV16, have been found in nearly all cases of cervical cancer. Therapies targeted at blocking the HPV16 E6 protein and its deleterious effects on the tumour suppressor pathways of the cell can reverse the malignant phenotype of affected keratinocytes while sparing uninfected cells. Through a strong interdisciplinary collaboration between engineering and biology, a novel, non-invasive intracellular delivery method for the HPV16 E6 antibody, F127-6G6, was developed. The method employs high intensity focused ultrasound (HIFU) in combination with microbubbles, in a process known as sonoporation. In this proof of principle study, it was first demonstrated that sonoporation antibody delivery into the HPV16 positive cervical carcinoma derived cell lines CaSki and SiHa was possible, using chemical transfection as a baseline for comparison. Delivery of the E6 antibody using sonoporation significantly restored p53 expression in these cells, indicating the antibody is able to enter the cells and remains active. This delivery method is targeted, non-cytotoxic, and non-invasive, making it more easily translatable for in vivo experiments than other transfection methods.

MatMRI and MatHIFU: software toolboxes for real-time monitoring and control of MR-guided HIFU

BackgroundThe availability of open and versatile software tools is a key feature to facilitate pre-clinical research for magnetic resonance imaging (MRI) and magnetic resonance-guided high-intensity focused ultrasound (MR-HIFU) and expedite clinical translation of diagnostic and therapeutic medical applications.In the present study, two customizable software tools that were developed at the Thunder Bay Regional Research Institute are presented for use with both MRI and MR-HIFU. Both tools operate in a MATLAB®; environment. The first tool is named MatMRI and enables real-time, dynamic acquisition of MR images with a Philips MRI scanner. The second tool is named MatHIFU and enables the execution and dynamic modification of user-defined treatment protocols with the Philips Sonalleve MR-HIFU therapy system to perform ultrasound exposures in MR-HIFU therapy applications.MethodsMatMRI requires four basic steps: initiate communication, subscribe to MRI data, query for new images, and unsubscribe. MatMRI can also pause/resume the imaging and perform real-time updates of the location and orientation of images. MatHIFU requires four basic steps: initiate communication, prepare treatment protocol, and execute treatment protocol. MatHIFU can monitor the state of execution and, if required, modify the protocol in real time.ResultsFour applications were developed to showcase the capabilities of MatMRI and MatHIFU to perform pre-clinical research. Firstly, MatMRI was integrated with an existing small animal MR-HIFU system (FUS Instruments, Toronto, Ontario, Canada) to provide real-time temperature measurements. Secondly, MatMRI was used to perform T2-based MR thermometry in the bone marrow. Thirdly, MatHIFU was used to automate acoustic hydrophone measurements on a per-element basis of the 256-element transducer of the Sonalleve system. Finally, MatMRI and MatHIFU were combined to produce and image a heating pattern that recreates the word ‘HIFU’ in a tissue-mimicking heating phantom.ConclusionsMatMRI and MatHIFU leverage existing MRI and MR-HIFU clinical platforms to facilitate pre-clinical research. MatMRI substantially simplifies the real-time acquisition and processing of MR data. MatHIFU facilitates the testing and characterization of new therapy applications using the Philips Sonalleve clinical MR-HIFU system. Under coordination with Philips Healthcare, both MatMRI and MatHIFU are intended to be freely available as open-source software packages to other research groups.

BerryPI: A software for studying polarization of crystalline solids with WIEN2k density functional all-electron package

Abstract We present a module that enables computation of polarization using density functional theory based on the full potential linearized augmented plane wave package WIEN2k . The theoretical background of deriving microscopic polarization of materials using the modern theory of polarization (geometric Berry phase approach) is reviewed. The software is validated and then applied to determine spontaneous polarization and Born effective charges of several crystal structures, which are commonly studied theoretically and experimentally.

Influence of cell line and cell cycle phase on sonoporation transfection efficiency in cervical carcinoma cells under the same physical conditions

Using cervical-carcinoma-derived cells as a model, the present study investigates the effects cell line and cell cycle phase have on sonoporation transfection efficiency under the same physical conditions. A plasmid expressing green fluorescent protein (GFP) was used to measure transfection efficiency. To evaluate the effect of cell type, CaSki, HeLa, and SiHa cells were sonoporated using an acoustic pressure of 1 MPa for 30 s with a duty cycle of 4.8% in the presence of the GFP plasmid. To study the effect of cell cycle phase, SiHa cells were synchronized at S-phase using a double thymidine block and sonoporated at different time points after the block. Contrast agent microbubbles were used at a 0.33% volume concentration. Results indicated that both cell line and cell cycle phase impact the transfection efficiency obtained with sonoporation.

Focused ultrasound treatment of abscesses induced by methicillin resistant Staphylococcus aureus: Feasibility study in a mouse model

PURPOSE To study the therapeutic effect of focused ultrasound on abscesses induced by methicillin-resistant Staphylococcus aureus (MRSA). MRSA is a major nosocomial pathogen where immunocompromised patients are prone to develop infections that are less and less responsive to regular treatments. Because of its capability to induce a rise of temperature at a very precise location, the use of focused ultrasound represents a considerable opportunity for therapy of localized MRSA-related infections. METHODS 50 μl of MRSA strain USA400 bacteria suspension at a concentration of 1.32 ± 0.5 × 10(5) colony forming units (cfu)/μl was injected subcutaneously in the left flank of BALB/c mice. An abscess of 6 ± 2 mm in diameter formed after 48 h. A transducer operating at 3 MHz with a focal length of 50 mm and diameter of 32 mm was used to treat the abscess. The focal point was positioned 2 mm under the skin at the abscess center. Forty-eight hours after injection four ultrasound exposures of 9 s each were applied to each abscess under magnetic resonance imaging guidance. Each exposure was followed by a 1 min pause. These parameters were based on preliminary experiments to ensure repetitive accurate heating of the abscess. Real-time estimation of change of temperature was done using water-proton resonance frequency and a communication toolbox (matMRI) developed inhouse. Three experimental groups of animals each were tested: control, moderate temperature (MT), and high temperature (HT). MT and HT groups reached, respectively, 52.3 ± 5.1 and 63.8 ± 7.5 °C at the end of exposure. Effectiveness of the treatment was assessed by evaluating the bacteria amount of the treated abscess 1 and 4 days after treatment. Myeloperoxidase (MPO) assay evaluating the neutrophil amount was performed to assess the local neutrophil recruitment and the white blood cell count was used to evaluate the systemic inflammatory response after focused ultrasound treatment. RESULTS Macroscopic evaluation of treated abscess indicated a diminution of external size of abscess 1 day after treatment. Treatment did not cause open wounds. The median (lower to upper quartile) bacterial count 1 day after treatment was 6.18 × 10(3) (0.76 × 10(3)-11.18 × 10(3)), 2.86 × 10(3) (1.22 × 10(3)-7.07 × 10(3)), and 3.52 × 10(3) (1.18 × 10(3)-6.72 × 10(3)) cfu/100 μl for control, MT and HT groups, respectively; for the 4-day end point, the count was 1.37 × 10(3) (0.67 × 10(3)-2.89 × 10(3)), 1.35 × 10(3) (0.09 × 10(3)-2.96 × 10(3)), and 0.07 × 10(3) (0.03 × 10(3)-0.36 × 10(3)) cfu/100 μl for control, MT and HT, showing a significant reduction (p = 0.002) on the bacterial load four days after focused ultrasound treatment when treating at high temperature (HT). The MPO amount remained unchanged between groups and days, indicating no change on local neutrophil recruitment in the abscess caused by the treatment. The white blood cell count remained unchanged between groups and days indicating that no systemic inflammatory response was caused by the treatment. CONCLUSIONS Focused ultrasound induces a therapeutic effect in abscesses induced by MRSA. This effect is observed as a reduction of the number bacteria without significantly altering the amount of MPO at the site of a MRSA-induced abscess. These initial results suggest that focused ultrasound is a viable option for the treatment of localized MRSA-related infections.

Multi‐frequency characterization of speed of sound for longitudinal transmission on freshly excised human skulls

The results of measurements of longitudinal speed of sound are presented for seven specimens of human calvaria. The study was done for frequencies between 0.27 and 2.525 MHz. Specimens were obtained from fresh cadavers through a protocol with the Division of Anatomy of the University of Toronto. The specimens were mounted in polycarbonate frames that were marked for stereoscopic positioning. CT scans of the skulls mounted on their frames were performed and a three‐dimensional reconstruction of the skull surface was done. A positioning system ensured normal sound incidence of an acoustic signal produced by a focused device with a diameter of 5 cm and a focal length of 10 cm. Speed of sound estimation was done with measurements of time‐of‐flight using a needle hydrophone (diameter of 0.5 mm) and a sound propagation model through layers that takes into account change of speed of sound in function of density. For six of seven specimens, measurements were done on five locations on the calvaria and for the othe...

Magnetic resonance-guided high-intensity focused ultrasound combined with radiotherapy for palliation of head and neck cancer-a pilot study.

BackgroundRadiotherapy is a critical component of the multidisciplinary management of cancers of the head and neck. It may comprise the primary curative treatment modality or is used in an adjuvant setting to improve local control and survival by preventing seeding and reseeding of distant metastases from persistent reservoirs of locoregional disease. Although considerable advances have been made recently in the fields of radiotherapy, systemic treatment and surgery for head and neck tumours, locoregional recurrence rates remain high and treatment side effects may have severe impact on patients’ quality of life.Magnetic resonance-guided high-intensity focused ultrasound (MRg-HIFU) is a novel technique in the treatment of cancer that has the potential to improve tumour cure rates and decrease treatment-related toxicity. Clinical applications of HIFU are being used increasingly for the treatment of several tumour sites, for example uterine leiomyomas and prostate cancer.Methods/DesignThe pilot study presented here is an initial step toward utilizing MRg-HIFU for head and neck cancer treatment. The rationale for novel treatment options in head and neck cancer is reviewed as well as emerging evidence that support the increasing clinical utilization of MRg-HIFU.DiscussionThis pilot study aims to assess safety, toxicity and feasibility of MRg-HIFU treatments to the head and neck region and to evaluate changes caused by MRg-HIFU within the treated tumour regions based on post-treatment MRI.

Class-DE Ultrasound Transducer Driver for HIFU Therapy

This paper presents a practical implementation of an integrated MRI-compatible CMOS amplifier capable of directly driving a piezoelectric ultrasound transducer suitable for high-intensity focused ultrasound (HIFU) therapy. The amplifier operates in Class DE mode without the need for an output matching network. The integrated amplifier has been implemented with the AMS AG H35 CMOS process. A class DE amplifier design methodology for driving unmatched piezoelectric loads is presented along with simulation and experimental results. The proposed design achieves approximately 90% efficiency with over 800 mW of output power at 1010 kHz. The total die area including pads is 2 mm2. Compatibility with MRI was validated with B1 imaging of a phantom and the amplifier circuit.