Stewart Russell

Gold Nanocrystal Labeling Allows Low-Density Lipoprotein Imaging from the Subcellular to Macroscopic Level

Low-density lipoprotein (LDL) plays a critical role in cholesterol transport and is closely linked to the progression of several diseases. This motivates the development of methods to study LDL behavior from the microscopic to whole-body level. We have developed an approach to efficiently load LDL with a range of diagnostically active nanocrystals or hydrophobic agents. We performed focused experiments on LDL labeled with gold nanocrystals (Au-LDL). The labeling procedure had minimal effect on LDL size, morphology, or composition. Biological function was found to be maintained from both in vitro and in vivo experiments. Tumor-bearing mice were injected intravenously with LDL, DiR-LDL, Au-LDL, or a gold-loaded nanoemulsion. LDL accumulation in the tumors was detected with whole-body imaging methods, such as computed tomography (CT), spectral CT, and fluorescence imaging. Cellular localization was studied with transmission electron microscopy and fluorescence techniques. This LDL labeling procedure should permit the study of lipoprotein biointeractions in unprecedented detail.

Security issues in the development of a wireless blood-glucose monitoring system

This paper is a progress report on an ongoing research effort that makes use of wireless biometric data management, specifically a wireless blood-glucose monitoring system (WBgM). The goal of this research is to realize appropriate timely intervention by health-care providers in response to records of unregulated blood-glucose level in order to maintain near-normal levels. The focus of this paper is security-their is the possibility that medical information may be used consciously by malicious eavesdropper to the detriment of a patient in search of new employment or a new insurer. There is the possibility of malicious or accidental data corruption from database intrusion. Mobile data transmission is especially problematic in preventing these. We present requirements and our approach taken to realize a secure system architecture. Key techniques and their implementation details to maintain privacy, authentication, and data integrity are discussed here.

Aquaporin-1 facilitates pressure-driven water flow across the aortic endothelium

Aquaporin-1, a ubiquitous water channel membrane protein, is a major contributor to cell membrane osmotic water permeability. Arteries are the physiological system where hydrostatic dominates osmotic pressure differences. In the present study, we show that the walls of large conduit arteries constitute the first example where hydrostatic pressure drives aquaporin-1-mediated transcellular/transendothelial flow. We studied cultured aortic endothelial cell monolayers and excised whole aortas of male Sprague-Dawley rats with intact and inhibited aquaporin-1 activity and with normal and knocked down aquaporin-1 expression. We subjected these systems to transmural hydrostatic pressure differences at zero osmotic pressure differences. Impaired aquaporin-1 endothelia consistently showed reduced engineering flow metrics (transendothelial water flux and hydraulic conductivity). In vitro experiments with tracers that only cross the endothelium paracellularly showed that changes in junctional transport cannot explain these reductions. Percent reductions in whole aortic wall hydraulic conductivity with either chemical blocking or knockdown of aquaporin-1 differed at low and high transmural pressures. This observation highlights how aquaporin-1 expression likely directly influences aortic wall mechanics by changing the critical transmural pressure at which its sparse subendothelial intima compresses. Such compression increases transwall flow resistance. Our endothelial and historic erythrocyte membrane aquaporin density estimates were consistent. In conclusion, aquaporin-1 significantly contributes to hydrostatic pressure-driven water transport across aortic endothelial monolayers, both in culture and in whole rat aortas. This transport, and parallel junctional flow, can dilute solutes that entered the wall paracellularly or through endothelial monolayer disruptions. Lower atherogenic precursor solute concentrations may slow their intimal entrainment kinetics.

Spatial frequency analysis of anisotropic drug transport in tumor samples

Abstract. Directional Fourier spatial frequency analysis was used on standard histological sections to identify salient directional bias in the spatial frequencies of stromal and epithelial patterns within tumor tissue. This directional bias is shown to be correlated to the pathway of reduced fluorescent tracer transport. Optical images of tumor specimens contain a complex distribution of randomly oriented aperiodic features used for neoplastic grading that varies with tumor type, size, and morphology. The internal organization of these patterns in frequency space is shown to provide a precise fingerprint of the extracellular matrix complexity, which is well known to be related to the movement of drugs and nanoparticles into the parenchyma, thereby identifying the characteristic spatial frequencies of regions that inhibit drug transport. The innovative computational methodology and tissue validation techniques presented here provide a tool for future investigation of drug and particle transport in tumor tissues, and could potentially be used a priori to identify barriers to transport, and to analyze real-time monitoring of transport with respect to therapeutic intervention.

Combination absorption and fluorescence for detection of vessels in intestinal sub-mucosa (Conference Presentation)

Injuries to main vascular structures within the sub mucosa present a serious complication during surgery. There is no evidence-based treatment to prevent this type of injury, so detection is critical. Using a combination of absorption and fluorescence imaging we can detect blood vessel phantoms to a depth of 7 mm in intestinal sub-mucosa. Using an illumination source at 850, and reading the cross-polarized reflected signal also at 850 gives the absorption image. Simultaneous excitation of ICG at 785 nm creates a fluorescent response that is used for contrast enhancement.

An interactive visual interface for the determination of similarity patterns in the Fourier spatial frequency spectrum of laser speckle

Laser speckle from particles that are smaller than the wavelength of light resemble a random Gaussian field, but can be shown to contain a characteristic spectrum in frequency space. Speckle is caused by not only the instantaneous microstructure of nanoparticles in suspension that will fluctuate as they reorganize, but also by the magnetic and optical properties of the scattering medium itself. Here we demonstrate interactive tool that can be used to define similarities between seemingly random scattering fields. Optimization of the Fourier spatial frequency spectrum gives a representative pattern that can be directly correlated to the transport properties of the particles.

Fourier spatial frequency analysis for image classification: training the training set

The Directional Fourier Spatial Frequencies (DFSF) of a 2D image can identify similarity in spatial patterns within groups of related images. A Support Vector Machine (SVM) can then be used to classify images if the inter-image variance of the FSF in the training set is bounded. However, if variation in FSF increases with training set size, accuracy may decrease as the size of the training set increases. This calls for a method to identify a set of training images from among the originals that can form a vector basis for the entire class. Applying the Cauchy product method we extract the DFSF spectrum from radiographs of osteoporotic bone, and use it as a matched filter set to eliminate noise and image specific frequencies, and demonstrate that selection of a subset of superclassifiers from within a set of training images improves SVM accuracy. Central to this challenge is that the size of the search space can become computationally prohibitive for all but the smallest training sets. We are investigating methods to reduce the search space to identify an optimal subset of basis training images.

Detection of particle flow patterns in tumor by directional spatial frequency analysis

Drug delivery to tumors is well known to be chaotic and limited, partly from dysfunctional vasculature, but also because of microscopic regional variations in composition. Modeling the of transport of nanoparticle therapeutics, therefore must include not only a description of vascular permeability, but also of the movement of the drug as suspended in tumor interstitial fluid (TIF) once it leaves the blood vessel. Understanding of this area is limited because we currently lack the tools and analytical methods to characterize it. We have previously shown that directional anisotropy of drug delivery can be detected using Directional Fourier Spatial Frequency (DFSF) Analysis. Here we extend this approach to generate flow line maps of nanoparticle transport in TIF relative to tumor ultrastructure, and show that features of tumor spatial heterogeneity can be identified that are directly related to local flow isometries. The identification of these regions of limited flow may be used as a metric for determining response to therapy, or for the optimization of adjuvant therapies such as radiation pre-treatment, or enzymatic degradation.

Directional spatial frequency analysis of lipid distribution in atherosclerotic plaque

Atherosclerosis is characterized by the growth of fibrous plaques due to the retention of cholesterol and lipids within the artery wall, which can lead to vessel occlusion and cardiac events. One way to evaluate arterial disease is to quantify the amount of lipid present in these plaques, since a higher disease burden is characterized by a higher concentration of lipid. Although therapeutic stimulation of reverse cholesterol transport to reduce cholesterol deposits in plaque has not produced significant results, this may be due to current image analysis methods which use averaging techniques to calculate the total amount of lipid in the plaque without regard to spatial distribution, thereby discarding information that may have significance in marking response to therapy. Here we use Directional Fourier Spatial Frequency (DFSF) analysis to generate a characteristic spatial frequency spectrum for atherosclerotic plaques from C57 Black 6 mice both treated and untreated with a cholesterol scavenging nanoparticle. We then use the Cauchy product of these spectra to classify the images with a support vector machine (SVM). Our results indicate that treated plaque can be distinguished from untreated plaque using this method, where no difference is seen using the spatial averaging method. This work has the potential to increase the effectiveness of current in-vivo methods of plaque detection that also use averaging methods, such as laser speckle imaging and Raman spectroscopy.

Characterization of the fourier spatial frequencies of time resolved laser speckle from interacting high density lipoprotein disks.

Fourier spatial frequency analysis of 2-D laser speckle images was used to identify the interaction of high density lipoprotein (HDL) nanoparticles in aqueous suspension at different ionic strengths. The decorrelation function of a time series of laser speckle images was used to identify key spectral ranges of HDL samples, and characteristic frequencies were extracted. These families of frequencies can be used as unique identifiers of nanometer scale structure in particle suspensions.