Improvements in Ultrasound Mediated Drug Delivery using Microbubble Contrast Agents and Application in Ex-Vivo Porcine Eyes

Ultrasound mediated drug delivery is a growing field which shows great promise. However, the ability to uniformly deliver acoustic energy to large 3D volumes of tissue remains a challenge for drug delivery. Additionally, uneven drug loading and large size distributions in microbubble contrast agents (MCAs) reduce the ability to control therapeutic dosing. The aim of this thesis is to demonstrate improvements in the field through new techniques in producing acoustically active drug delivery agents and applying ultrasonic parameters to volumetric regions of interest. The use of a computer controlled motion stage to apply ultrasound to a 3D volume of tissue is shown, allowing the uniform delivery of destructive pulses and acoustic radiation force. This technique is demonstrated in a novel application for treating diseases of the posterior segment of the eye. Injections of MCAs loaded with fluorescent dye into the suprachoroidal space of ex-vivo porcine globes are used to assess the feasibility of ultrasound mediated drug delivery for treating tissues in the rear of the eye. Also demonstrated is the production of MCAs with a controlled size and drug loading distribution for oil soluble compounds, which could enable greater control over therapeutic dosing.Master of Scienc

Space Station Freedom/lunar transfer vehicle propellant operation hazard analysis

Space Station Freedom (SSF), as a transportation node for Space Exploration Initiative missions, would involve the assembly and refurbishing of lunar and Mars transfer vehicles. This includes operations involving cryogenic propellants (LH2 7 LO2) such as storing and handling of loaded propellant tanks, assembly onto the vehicle, and propellant transfer. Cryogenic propellants dictate rigorous safety precautions and impose unique requirements to ensure flight safety to both personnel and SSF elements. The objective of this study is to identify potential hazards and risks associated with cryogenic propellants. This involves identification of pertinent system design features and operational procedures. Criticality of identified risks/hazards shall be assessed and those that fall in the catastrophic and critical categories shall include mitigating solutions

Effect and Distribution of Contrast Medium after Injection into the Anterior Suprachoroidal Space in Ex Vivo Eyes

PURPOSE: To determine the effects and posterior distribution of injections made into the anterior suprachoroidal space (SCS). METHODS: The anterior SCS of adult porcine and canine ex vivo eyes was cannulated. Latex injections and high frequency ultrasound (50 MHz) was used to image the effect and distension of the SCS. Flow characteristics and percentage maximal distribution of microbubble contrast injection into the SCS were assessed by 2D and 3D ultrasound. RESULTS: Mean (SD) distension of the SCS with PBS increased from 1.57 (0.48) mm after injection of 250 μL to 3.28 (0.57) mm with 1000 μL PBS. Eyes injected at physiologic IOP had no significant difference in SCS distension. In real-time 2D ultrasound, the contrast agent flowed from the injection site to the opposite ventral anterior SCS and the posterior SCS. Contrast arrived at the opposite and posterior SCS 7.8 (4.6) and 7.7 (4.6) seconds after injection, respectively. In sagittal images, contrast was visible in 24.0%to 27.2% of the SCS; in 10 of 12 eyes, contrast reached the posterior pole of the eye. In 3D images, contrast medium occupied 39.0% to 52.1% of the entire SCS. CONCLUSIONS: These results suggest that the SCS can expand, in a dose-dependent manner, to accommodate various volumes of fluid and that it is possible to image the SCS with ultrasound contrast. The authors' hypothesis that a single anterior SCS injection can reach the ocular posterior segment was supported. Further development of SCS injections for treatment of the ocular posterior segment is warranted

Effects of Cloud Horizontal Inhomogeneity and Drizzle on Remote Sensing of Cloud Droplet Effective Radius: Case Studies Based on Large-eddy Simulations

This study investigates effects of drizzle and cloud horizontal inhomogeneity on cloud effective radius (re) retrievals from the Moderate Resolution Imaging Spectroradiometer (MODIS). In order to identify the relative importance of various factors, we developed a MODIS cloud property retrieval simulator based on the combination of large-eddy simulations (LES) and radiative transfer computations. The case studies based on synthetic LES cloud fields indicate that at high spatial resolution (100 m) 3-D radiative transfer effects, such as illumination and shadowing, can induce significant differences between retrievals ofre based on reflectance at 2.1 m (re,2.1) and 3.7 m (re,3.7). It is also found that 3-D effects tend to have stronger impact onre,2.1 than re,3.7, leading to positive difference between the two (re,3.72.1) from illumination and negative re,3.72.1from shadowing. The cancellation of opposing 3-D effects leads to overall reasonable agreement betweenre,2.1 and re,3.7 at high spatial resolution as far as domain averages are concerned. At resolutions similar to MODIS, however, re,2.1 is systematically larger than re,3.7when averaged over the LES domain, with the difference exhibiting a threshold-like dependence on bothre,2.1and an index of the sub-pixel variability in reflectance (H), consistent with MODIS observations. In the LES cases studied, drizzle does not strongly impact reretrievals at either wavelength. It is also found that opposing 3-D radiative transfer effects partly cancel each other when cloud reflectance is aggregated from high spatial resolution to MODIS resolution, resulting in a weaker net impact of 3-D radiative effects onre retrievals. The large difference at MODIS resolution between re,3.7 and re,2.1 for highly inhomogeneous pixels with H 0.4 can be largely attributed to what we refer to as the plane-parallelrebias, which is attributable to the impact of sub-pixel level horizontal variability of cloud optical thickness onre retrievals and is greater for re,2.1 than re,3.7. These results suggest that there are substantial uncertainties attributable to 3-D radiative effects and plane-parallelre bias in the MODIS re,2.1retrievals for pixels with strong sub-pixel scale variability, and theH index can be used to identify these uncertainties

Effect of Essential Fatty Acid Deficiency on Cutaneous Sterol Synthesis

The fact that the skin is a major site of total body sterologensis, coupled both with the apparent absence of low density lipoprotein receptors on keratinocytes and with the lack of influence of serum cholesterol on epidermal sterologensis, has created the impression that epidermal lipid synthesis might be autonomous, i.e. nonregulatable. Recent studies have shown, however, that disruption of cutaneous barrier function with acetone or detergents stimulates epidermal sterologensis (J Lipid Res 26:418–427, 1985). To correlate further sterologenesis with barrier function, we measured de novo synthesis of cholesterol and total nonsaponifiable lipids in essential fatty acid-deficient (EFAD) hairless mice. Animals with defective barrier function, manifested by abnormal transepidermal water loss, demonstrated a 2-fold increase in epidermal cholesterol and total nonsaponifiable lipid synthesis over controls while synthesis in the dermis was unchanged. Epidermal sterologenesis in EFAd animals, repleted with linoleic acid either systematically or topically, returned toward normal as barrier function improved. Moreover, plastic occlusion of EFAD mouse skin normalized epidermal sterologensis at 1 and 3 days. These results provide further evidence that epidermal sterologenesis is not entirely autonomous, and can be regulated by water barrier requirements

The Atmospheric Radiation Measurement Program May 2003 Intensive Operations Period Examining Aerosol Properties and Radiative Influences: Preface to Special Section

Atmospheric aerosols influence climate by scattering and absorbing radiation in clear air (direct effects) and by serving as cloud condensation nuclei, modifying the microphysical properties of clouds, influencing radiation and precipitation development (indirect effects). Much of present uncertainty in forcing of climate change is due to uncertainty in the relations between aerosol microphysical and optical properties and their radiative influences (direct effects) and between microphysical properties and their ability to serve as cloud condensation nuclei at given supersaturations (indirect effects). This paper introduces a special section that reports on a field campaign conducted at the Department of Energy Atmospheric Radiation Measurement site in North Central Oklahoma in May, 2003, examining these relations using in situ airborne measurements and surface-, airborne-, and space-based remote sensing

3-D Microvessel-Mimicking Ultrasound Phantoms Produced With a Scanning Motion System

Ultrasound techniques are currently being developed which can assess the vascularization of tissue as a marker for therapeutic response. Some of these ultrasound imaging techniques seek to extract quantitative features about vessel networks, while high-frequency imaging also allows individual vessels to be resolved. The development of these new techniques, and subsequent imaging analysis strategies, necessitates an understanding of their sensitivities to vessel and vessel network structural abnormalities. Constructing in-vitro flow phantoms for this purpose can be prohibitively challenging, as simulating precise flow environments with non-trivial structures is often impossible using conventional methods of construction for flow phantoms. Presented in this manuscript is a method to create predefined structures with < 10 μm precision using a three-axis motion system. The application of this technique is demonstrated for the creation of individual vessel and vessel networks, which can easily be made to simulate the development of structural abnormalities typical of diseased vasculature in-vivo. Additionally, beyond facilitating the creation of phantoms which would be otherwise very challenging to construct, the method presented herein enables one to precisely simulate very slow blood flow, respiration artifacts, and to measure imaging resolution

Quantitative Volumetric Perfusion Mapping of the Microvasculature Using Contrast Ultrasound

Contrast-enhanced ultrasound imaging has demonstrated significant potential as a noninvasive technology for monitoring blood flow in the microvasculature. With the application of nondestructive contrast imaging pulse sequences combined with a clearance-refill approach, it is possible to create quantitative time-to-refill maps of tissue correlating to blood perfusion rate. One limitation to standard two-dimensional (2D) perfusion imaging is that the narrow elevational beamwidth of 1- or 1.5-D ultrasound transducers provides information in only a single slice of tissue, and thus it is difficult to image exactly the same plane from study to study. We hypothesize that inhomogeneity in vascularization, such as that common in many types of tumors, makes serial perfusion estimates inconsistent unless the same region can be imaged repeatedly. Our objective was to evaluate error in 2D quantitative perfusion estimation in an in vivo sample volume because of differences in transducer positioning. To mitigate observed errors due to imaging plane misalignment, we propose and demonstrate the application of quantitative 3-dimensional (3D) perfusion imaging. We also evaluate the effect of contrast agent concentration and infusion rate on perfusion estimates. Contrast-enhanced destruction-reperfusion imaging was performed using parametric mapping of refill times and custom software for image alignment to compensate for tissue motion. Imaging was performed in rats using a Siemens Sequoia 512 imaging system with a 15L8 transducer. A custom 3D perfusion mapping system was designed by incorporating a computer-controlled positioning system to move the transducer in the elevational direction, and the Sequoia was interfaced to the motion system for timing of the destruction-reperfusion sequence and data acquisition. Perfusion estimates were acquired from rat kidneys as a function of imaging plane and in response to the vasoactive drug dopamine. Our results indicate that perfusion estimates generated by 2D imaging in the rat kidney have mean standard deviations on the order of 10%, and as high as 22%, because of differences in initial transducer position. This difference was larger than changes in kidney perfusion induced by dopamine. With application of 3D perfusion mapping, repeatability in perfusion estimated in the kidney is reduced to a standard deviation of less than 3%, despite random initial transducer positioning. Varying contrast agent administration rate was also observed to bias measured perfusion time, especially at low concentrations; however, we observed that contrast administration rates between 2.7 × 108 and 3.9 × 108 bubbles/min provided results that were consistent within 3% for the contrast agent type evaluated. Three-dimensional perfusion imaging allows a significant reduction in the error caused by transducer positioning, and significantly improves the reliability of quantitative perfusion time estimates in a rat kidney model. When performing perfusion imaging, it is important to use appropriate and consistent contrast agent infusion rates to avoid bias

Realism of Lagrangian Large Eddy Simulations Driven by Renalysis Meteorology: Tracking a Pocket of Open Cells Under a Biomass Burning Aerosol Layer

An approach to drive Lagrangian large eddy simulation (LES) of boundary layer clouds with reanalysis data is presented and evaluated using satellite (Spinning Enhanced Visible and Infrared Imager, SEVIRI) and aircraft (Cloud‐Aerosol‐Radiation Interactions and Forcing, CLARIFY) measurements. The simulations follow trajectories of the boundary layer flow. They track the formation and evolution of a pocket of open cells (POC) underneath a biomass burning aerosol layer in the free troposphere. The simulations reproduce the evolution of observed stratocumulus cloud morphology, cloud optical depth, and cloud drop effective radius, and capture the timing of the cloud state transition from closed to open cells seen in the satellite imagery on the three considered trajectories. They reproduce a biomass burning aerosol layer identified by the in‐situ aircraft measurements above the inversion of the POC. Entrainment of aerosol from the biomass burning layer into the POC is limited to the extent of having no impact on cloud‐ or boundary layer properties, in agreement with the CLARIFY observations. The two‐moment bin microphysics scheme used in the simulations reproduces the in‐situ cloud microphysical properties reasonably well. A two‐moment bulk microphysics scheme reproduces the satellite observations in the non‐precipitating closed‐cell state, but overestimates liquid water path and cloud optical depth in the precipitating open‐cell state due to insufficient surface precipitation. A boundary layer cold and dry bias occurring in LES can be counteracted by reducing the grid aspect ratio and by tightening the large scale wind speed nudging towards the surface. </div

Controllable microfluidic synthesis of multiphase drug-carrying lipospheres for site-targeted therapy

We report the production of micrometer-sized gas-filled lipospheres using digital microfluidics technology for chemotherapeutic drug delivery. Advantages of on-chip synthesis include a monodisperse size distribution (polydispersity index (σ) values of <5%) with consistent stability and uniform drug loading. Photolithography techniques are applied to fabricate novel PDMS-based microfluidic devices that feature a combined dual hydrodynamic flow-focusing region and expanding nozzle geometry with a narrow orifice. Spherical vehicles are formed through flow-focusing by the self-assembly of phospholipids to a lipid layer around the gas core, followed by a shear-induced break off at the orifice. The encapsulation of an extra oil layer between the outer lipid shell and inner bubble gaseous core allows the safe transport of highly hydrophobic and toxic drugs at high concentrations. Doxorubicin (Dox) entrapment is estimated at 15 mg mL−1 of particles packed in a single ordered layer. In addition, the attachment of targeting ligands to the lipid shell allows for direct vehicle binding to cancer cells. Preliminary acoustic studies of these monodisperse gas lipospheres reveal a highly uniform echo correlation of greater than 95%. The potential exists for localized drug concentration and release with ultrasound energy