Appendix A: Strategy for Nanotechnology-related EHS Research

Limitations of the data provided in this appendix.

This table of nanotechnology EHS projects provide the following information for each project:

Award # 2R01DK047858-10A1 Agency NIH Lead Institution University of FloridaTypeExtramural

Explanatory Notes This project will pave the way for further imaging studies in tissue engineering that may be applicable to the nanoscale.

Abstract The main criterion in assessing the therapeutic efficacy of tissue engineered construct is the successful restoration of the host's physiology. Direct and non-invasive in vivo monitoring of a construct is of great importance for the following reasons: it provides correlations between construct function and physiologic effects post-implantation in vivo; and it offers the possibility of assessing changes in construct function that may be used to develop early markers of construct failure in advance of the end-point effects. The overall objective of the proposed research is to develop a noninvasive methodology to monitor in vivo the function of an implanted tissue engineered pancreatic substitute. NMR is uniquely suited to perform such studies since it is a non-invasive modality that has the ability to probe into the intracellular metabolism of the construct, and to view the anatomy of the host at the site of implantation. NMR spectroscopic and imaging techniques can monitor several intracellular and extracellular metabolites without necessitating the introduction of foreign agents or the fixation of cells. It is our hypothesis that NMR detectable metabolic indexes can be used to monitor the function of an implanted tissue engineered pancreatic substitute and provide early indicators of implant failure while the recipient is still euglycemic. The NMR nuclei examined in this study include 1H, 19F and 31P, while the model pancreatic construct utilized is composed of mouse insulinoma #TC-tet cells or mouse islets encapsulated in alginate/poly-L- lysine/alginate beads and contained within an agarose matrix that allows for easy retrieval. The following Specific Aims are designed to address our hypothesis: Specific Aim 1: To optimize in vivo NMR signal acquisition with inductively coupled RF-coils. Specific Aim 2: To develop and validate a model of time-dependent oxygen and cell density gradients within constructs. Specific Aim 3: To non-invasively assess the function of an implanted tissue engineered pancreatic substitute and correlate that to end point physiologic events. We believe this to be a thorough, quantitative and a significant study to monitor the function of an implanted tissue engineered substitute that will identify metabolic events associated with failure ahead of its end physiologic effect. Proposed experiments are founded on strong preliminary data and we are confident that they will pave the way for further imaging studies in tissue engineering.

Award # 1Z01HD000261-09 Agency NIH Lead Institution NICHD/SBSPTypeIntramural

Explanatory Notes A critical element of this project is the development of methods and instrumentation to image tissues and study biological phenomena with nanoscale resolution.

Abstract Scientists in the Section on Biomedical Stochastic Physics (SBSP) devise quantitative theories, develop methodologies, and design instrumentation to study biological phenomena whose properties are characterized by elements of randomness in both space and time. The research focuses on developing quantitative theories applicable to quantitative optical spectroscopy and tomographic imaging of tissues. This requires analyzing different optical sources of contrast such as endogenous or exogenous fluorescent labels, absorption (e.g., hemoglobin or chromophore concentration), and/or scattering. SBSP researchers design and conduct experiments and computer simulations to validate theoretical findings. In addition, collaborations formed with other scientists at the NIH and researchers around the country and world investigate physiological sites where optical techniques might be clinically practical and offer new diagnostic knowledge and/or less morbidity than existing diagnostic methods. Biological tissues often exhibit characteristic regular features or ornamental patterns. Transition from normal tissue function to diseased tissue can be detected by quantifying irregular patterns. The degree of statistical similarities in a region of interest can carry valuable comparative information about the structural features of the tissue and can help to characterize tissue, i.e., analyze disease localization and progression. To visualize subsurface structural features of biological tissues, we have developed a user-friendly polarization imaging system that simultaneously images cross- and co-polarized light. We have developed a quantitative statistical tool, based on Pearson correlation coefficient analysis to enhance the image quality and reveal regions of high statistical similarities within the noisy tissue images. We have shown that under certain conditions, such maps of the correlation coefficient are determined by the textural character of tissues and not the choice of the reference image region, providing information on tissue structure. As an example, the subsurface texture of a demineralized tooth sample was enhanced from a noisy polarized light image. Many biological tissues (muscle, skin, white matter in brain, etc.) are known to be anisotropic, i.e., photons tend to migrate preferentially along fibers. To consider the effects of tissue anisotropy on observed characteristics of fluorescent light, we have generalized our random walk analysis of light propagation in the anisotropic turbid media for the case of a deeply embedded small fluorophore or scattering inclusion with special focus on the time-resolved measurement set-up. Our goal is to find an analytical expression for the expected change in the photon mean time of flight due to the presence of such an abnormality. Fluorophore lifetime imaging is a promising tool for studying tissue environment such as tumors. The lifetime (time for an electron to return from excited state to initial state) of a fluorophore can vary in response to changes in the immediate environment such as temperature, pH, tissue oxygen content, nutrient supply, and bioenergetic status. Mapping the lifetime and location of a fluorophore in tissue at different depths can be used to monitor such parameters. Toward this goal, we have developed a time-resolved lifetime imaging system for in vivo small animal studies that maps fluorophores lifetimes. The system consists of a single source-multiple detector array that scans the surface of the tissue. Using several source-detector separations, one is able to probe different depths of the medium. In collaboration with Dr. Capala in the Radiation Oncology Program of NCI who has developed a pH sensitive dye in the near-infrared region, we have studied the tumor environment below the skin. We have demonstrated that by using simplified back projections we are able to map near surface fluorescent lifetime in vivo. Combining this with the pre-calibrated lifetime response to pH, we have shown that biologically plausible, non-invasive, quantification of pH in mouse tumors can be determined. The oncology community is testing a number of novel targeted approaches for use against a variety of cancers. With regard to monitoring vasculature, it is desirable to develop and assess noninvasive and quantitative techniques that can not only monitor structural changes, but can also assess the functional characteristics or the metabolic status of the tumor. We are testing three potential noninvasive imaging techniques to monitor patients undergoing an experimental therapy: infrared thermal imaging (thermography), laser Doppler imaging (LDI) and multi-spectral imaging. These imaging techniques are being tested on subjects with Kaposis sarcoma (KS), a highly vascular tumor that occurs frequently among people infected with acquired immunodeficiency syndrome (AIDS). Cutaneous KS lesions are easily accessible for noninvasive techniques that involve imaging of tumor vasculature, and they thus represent a tumor model in which to assess certain parameters of angiogenesis. The KS studies are ongoing clinical trials under four different NCI protocols. Thermography graphically depicts temperature gradients over a given body surface area at a given time. LDI can more directly measure the net blood velocity of small blood vessels in tissue, which generally increases as blood supply increases during angiogenesis. NIRS is most closely related to visual assessment. In collaboration with Dr. Demos at the Lawrence Livermore National Laboratory, a portable spectral imaging system was designed that captures images with a high-resolution CCD camera at six near-infrared wavelengths (700, 750, 800, 850, 900, and 1000 nm). Collected intensity images are used in a mathematical optical model of skin containing two layers: an epidermis and much thicker, highly scattering dermis. Each layer contains major chromophores that determine absorption in the corresponding layer and the layers together determine the total reflectance of the skin. Local variations in melanin, oxygenated hemoglobin (HbO2), and blood volume are reconstructed through a multivariate analysis. High-resolution confocal laser microscopy is an intensively active field in modern bioimaging technologies because this technique provides sharp, high-magnification, three dimensional imaging with submicron resolution by non-invasive optical sectioning and rejection of out-of-focus information. We have developed a simple fiber-optic confocal microscope with nanoscale depth resolution beyond the diffraction barrier. It is based on combining the advanced properties of a simple apertureless single-mode-fiber confocal microscope design that provides highly sensitive diffraction-free Gaussian point light source/receiver, and a differential confocal microscope approach in which the sharp diffraction free slope of the axial confocal response curve is exploited. We have also developed an algorithm to enhance diffraction-limited images and obtain information on features smaller than the diffraction limit. Our algorithm tries to infer the best estimate of an object based on the diffraction-limited input image. Imaging an object with a diffraction-limited lens introduces in a blurred image, where neighboring pixels on the camera are correlated. The correlations between pixels are determined by the point spread function (PSF) of a diffraction-limited lens.

Award # 1R01MH071830-01A2 Agency NIH Lead Institution Washington State UniversityTypeExtramural

Explanatory Notes In combination with other intracellular measurement techniques, which may involve nanoparticles like quantum dots, this technique will improve ways of correlating electrophysiological measurements with intracellular biochemical or genetic pathway measurements not currently done.

Abstract The impact of sleep loss and sleep disorders on the health, social and economic well being of Americans is enormous. Yet our knowledge about the control and function of sleep remains severely limited, and based largely on studies where subjects are tethered, with significant behavioral side- effects. Thus, compact, implantable recording systems have become a major factor in sleep studies, especially in small transgenic mouse models where tethering is not practical. Existing telemetry systems are severely limited in the amount of information they can gather, and are not conducive for most studies. To address this need, we have assembled an interdisciplinary team to address four specific aims. First, we will develop a flexible electrode array that can be chronically implanted on the cortical surface of neural tissue. Traditional rigid electrode arrays require large skull openings. The flexible array has the advantage of minimal tissue trauma because only a slot in the skull is needed to insert the flexible array. One major drawback of multi-channel recordings comes from the large number of wires required for 16 to 256 or more channels of electrophysiology. Signals multiplexing can help, but available components including amplifiers, filters and multiplexers are comparatively large. Thus, our second aim will develop a miniature analog-system-on-a-chip, including preamplifiers, filters, multiplexer and 16 bit analog-to-digital converter for sampling 16 to 256 channels up to 32 kHz per channel. The chip will initially require only 5 wires for a serial digital connection and will weigh less than 1 gram. Our third aim will be to implement high speed wireless technology to allow the serial digital data to be transmitted directly from the acquisition chip to our computer interface card without the use of wires. Recent developments in digital wireless technology have allowed unprecedented data rates through transmitted signals. However, in order for wireless technology to be effective, our fourth aim will focus on an implantable power source enabling fully untethered recordings. Different power source technologies will be explored including battery and magnetic inductance. The technologies developed within this proposal will provide powerful new tools for neuroscience when many channels of electrophysiology, EEC, SEP and multi-unit electrodes are required in freely behaving animals, especially small rodents. The new technology is particularly important for wireless medical devices that require many channels with high data rates.

Award # 1R21MH074703-01A1 Agency NIH Lead Institution Johns Hopkins UniversityTypeExtramural

Explanatory Notes This research is in neural micro/nano systems within a biomedical instrumentation laboratory developing technologies for recording from neurons or the brain and developing interfaces, at molecular/cellular and at the systems level. Micro- and nanotechnology are utilized for the fabrication of sensors.

Abstract This is an interdisciplinary, innovative technology research and demonstration project to develop the next generation of neural probes for implanted recording. Current neural probes record electrical signals with the use of multiple silicon micromachined electrodes. In an advancement in this field, supported by previous NIH funding, we have demonstrated miniature neurochemical sensors and developed the specialized VLSI integrated circuitry needed to enable chemical measurements from multiple electrodes in a microprobe assembly. The next challenge is to research methods to harvest power to energize the implanted sensor and circuitry. Towards this goal, we propose two innovative technologies 1) the development of a rechargeable, microbattery system capable of sustaining power to the sensor and circuitry, 2) development of a novel VLSI wireless power harvesting circuit to energize the battery. Our other aims are to 3) develop an integrated probe with neurochemical sensor, VLSI wireless interface, power harvesting circuit and microbattery, and 4) evaluate the probe in a model of global ischemic brain injury. This research contributes to our long term goal to build fully implantable, autonomous microprobes, without any tethering, for neurochemical recording in chronically instrumented and tether less animals. A fully self-powered implanted neural microprobe system will be an enabling tool in the hands of neuroscientists interested in recording neural activity from animal models of brain function or brain disorders with the use of microelectrode arrays.

Award # 1R21MH078822-01 Agency NIH Lead Institution University of ChicagoTypeExtramural

Explanatory Notes This work supports nano- to micro-scale fluorescence measurements of electric fields in molecules.

Abstract The long term objective of this project is the development of an optical detection system based on surface plasmon resonance to study the dynamics of membrane proteins with special emphasis in voltage gated ion channels such as the Na and K channels that are responsible for the generation and propagation of the nerve impulse. Channel proteins are labeled in specific sites with fluorescent probes using cysteine chemistry and the fluorescence is detected by the proposed optical setup. Fluorescence changes, produced by quenching or energy transfer are indicators of local environmental changes and thus they follow conformational changes within the protein as the channel undergoes transitions from the closed to the open state. The optical apparatus uses a hemispherical lens that couples an incoming laser beam on a glass chip that has a thin (50 nm) siver layer where the biological preparation lies separated by a thin (10 nm) layer of silicon oxide. The correct angle of excitation induces plasmon resonance in the metal and enhances the fluorescence of fluorophores labeling the channel. The detection is done from the biological preparation side or from the excitation side. In the second case the signal to noise ratio is expected to be much larger because the coupled emission comes from a region limited to 20 nm and, as it is directional, a specially designed optics collects most of the light on a photodetector. The testing of the optical sytem is done on labeled ion channels expressed in mammalian cells or in supported bilayers. In the second case, the supported bilayer is made with liposomes containing purified labeled channels. The voltage across the bilayer is changed taking advantage of the silver layer of the plasmon chip. A modification of the optical system is also proposed to image the biological preparation to follow the time course of the fluorescence of individual molecules in response to voltage pulses that change the conformation of the channel. The understanding of conformational dynamics of channel proteins is a crucial step in the design of drugs or therapies needed to ameliorate or cure several neurological deseases produced by abnormal function of ion channels. The optical system developed in this application is aimed at developing a new microscope that is especially designed to detect conformational changes of ion channels with improved resolution, higher sensitivity and improved rejection of spurious fluorescence than presently available devices.

Award # 1R43MH076318-01 Agency NIH Lead Institution Pinnacle Technology, Inc.TypeExtramural

Explanatory Notes In combination with other intracellular measurement techniques, which may involve nanoparticles like quantum dots, this technique will improve ways of correlating electrophysiological measurements with intracellular biochemical or genetic pathway measurements in ways not currently done.

Abstract The long term objective of this project is to develop, validate and commercialize wireless, head mounted, turnkey, EEG/EMG systems with an integrated biosensor for rats and mice. A further objective is to develop a 76 uM glucose biosensor for direct measurement of brain glucose levels in rats and mice. The specific aims of this Phase I project are to develop and test a 76 uM glucose biosensor, a tethered EEG/EMG/Biosensor solution for mice and a wireless solution for rats. To reach these objectives, Pinnacle Technology, Northwestern University and the University of Kansas are building on past successes in the design of glutamate biosensors, wireless potentiostats for rats, and tethered EEG/EMG systems for mice. These products were developed in separate collaborations and are currently being commercialized. Products to be introduced include: a tethered system for mice, a wireless system for rats, and ultimately a fully wireless system for mice. Commercial applications include sleep research, behavioral research and drug screening. Technological innovations include biosensor design, turnkey head mount design, advanced electronics design, and advanced low power radio frequency design. The ability to measure glucose from specific brain areas in vivo while simultaneously recording sleep in rodents will give researchers the ability to better examine the functioning of specific sites within the brain during the sleep process as well as leverage the advantages conferred by using rat and mouse models for research. At the moment, it is not possible to concurrently study sleep and glucose regulation in a mouse model and there is only one published account where it has been attempted in the rat. The ability to instantly record glucose levels in a sleeping mouse or rat, and correlate that activity with EEG/EMG, will be valuable to researchers studying sleep and metabolism. The investigation of glucose in a mouse model will open up new avenues of research with genetic mutations available in species such as the NIRKO mouse (brain/neuron-specific insulin receptor knockout) which may have altered glucose responses during sleep and provide clues as to how sleep and metabolism are linked.

Award # 5R21EB005390-02 Agency NIH Lead Institution TDA Research, Inc.TypeExtramural

Explanatory Notes This project strives to improve magnetic resonance techniques for cellular and molecular imaging through the development of new contrast agents based on nanotubes and magnetic nanomaterials. Biocompatibility and physical characterization studies are central components.

Abstract This project will investigate a new class of potent magnetic resonance imaging (MRI) contrast agents based on cut carbon nanotubes filled with magnetic nanomaterials. These materials will function as protective nanocapsules for the delivery of magnetic materials and offer orders-of-magnitude improvement in MR contrast enhancement, vital to achieving the goal of cellular and molecular imaging with MRI. Synthetic efforts towards filling the cut nanotubes and derivatizing their outer surfaces for water solubility and biocompatibility will be central. Physical characterization of this new class of MRI contrast agent, including relaxometry by NMRD will provide insight into their relaxation mechanisms and offer avenues for further improvement. Collaboration with experts in toxicology and radiology will establish their suitability for eventual clinical investigations. The new platform technology to be created in this project offers a unique paradigm in MRI contrast enhancement useful for broad applications as both an implement in the medical research toolkit as well as for clinically relevant MRI contrast agents for the detection and improved treatment of disease.

Award # 5R21EB005365-02 Agency NIH Lead Institution Sandia National LaboratoryTypeExtramural

Explanatory Notes This effort to optimize the design, delivery, and imaging of the novel nanoprobes will lead to the development and improvement of methods to detect and characterize nanomaterials.

Abstract The major goal of this application is to synthesize new ceramic nanoprobes for bio-imaging that are highly fluorescent, bio-compatible, non-toxic, and tunable. This will be accomplished through a cross-discipinary venture between: (i) a group of material scientists and chemists from Sandia National Laboratories (Sandia) who are new to the NIH but have proven expertise in nanoparticle synthesis and biofurctionalization and (ii) a group of NIH-funded cell biologists from the University of New Mexico's School of Medicine (UNM-SOM) who have expertise in imaging cell signaling and trafficking pathways. The Sandia chemistry group will work concurrently towards two goals: (i) the synthesis of luminescent ceramic nanoprobes and (ii) innovations in functionalization that will deliver these probes to desired cellular targets. Initial synthetic efforts will focus on generating analogs of naturally occurring fluorescent (NOF) minerals as well as developing tailor-made nanoparticles doped with lanthanide cations for enhanced fluorescence. A library of novel compounds and synthetic pathways previously developed in the Sandia laboratory for nanoparticle synthesis will provide a unique knowledge base to initiate the development of useful luminescent ceramic nanoprobes. Probe functionalization will rely on PEG-phospholipids that allow for further bio-conjugation with proteins, peptides, small organic molecules/ligands and also with poly(arginine)-based transporters for transmembrane delivery. The UNM biology group will provide bioma terials to assist in the functionalization of the nanoprobes and will image probe delivery and specificity for cellular targets using live cell multispectral confocal microscopy. Preliminary interdisciplinary studies have validated the basic approaches for the synthesis of NOF nanoprobes and the biodelivery and imaging of nanoparticles. Intensive work to optimize the design, delivery, and imaging of these new nanoprobes is expected to achieve the RFA's goal of "increases in the sensitivity and specificity of molecular probes" for imaging. Results from this award will set the stage for in vivo studies whose goals will be: (i) to detect cancer sites, inflammation, and other disease processes and (ii) to deliver and release drugs at disease sites by further manipulation of the functionalization chemistry. All of the innovations uncovered during the preparation of these nanoprobes will be shared in order to benefit medical research applications beyond those proposed in this investigation.

Award # 5R21EB005364-02 Agency NIH Lead Institution Montana State UniversityTypeExtramural

Explanatory Notes This project aims to achieve a one-order-of-magnitude increase in the ability to detect and image molecular level events in vitro and in vivo. Improvement to techniques for detecting nano-particle contrast agents supports metrology methods to detect nanomaterials.

Abstract Our overall goal is to create and test a new generation of cell- targeted MRI contrast agents. The aim will be to achieve at least an order of magnitude increase in improvement in the ability to detect and image molecular level events in vitro and in vivo with broad applications in medicine. This proposal is a multidisciplinary effort, combining established expertise in cardiovascular, vascular biology, MRI, virology, synthetic inorganic and nano-materials chemistry. The overall approach will combine the use of non-infectious virus and other protein cage architectures for multivalent display of cell-specific targeting ligands, high performance metal based imaging agents, for functional and cellular imaging using MRI. The advantage of this approach is a substantial increase in rotational correlation time due to the size of the virus architecture in combination with high-density presentation of targeting ligands and metal binding sites with free access to water. The specific objectives of this proposal are (1) synthesis of protein cage architectures with high-density presentation of both cell targeting ligands and high magnetic moment materials, (ii) controlled fabrication of protein cage nano-particle clusters incorporating functionalized cell targeting ligands and MRI contrast agents, and (Hi) in vitro and in vivo MRI evaluation of functionalized protein cage architectures including improved MRI techniques for detecting nano-particle contrast agents. Creation and evaluation of these materials as functional MR contrast agents using state-of the-art facilities will provide rapid and direct feedback for an iterative process to create the next generation of high performance functional MRI contrast agents.

Award # 5R21EB005394-02 Agency NIH Lead Institution University of Texas-MD AndersonTypeExtramural

Explanatory Notes This project targets the development of ultra-sensitive magnetic resonance imaging probes for cancer molecular imaging applications. To evaluate the safety, biocompatibility, and effectiveness of probe materials (which include nanostructured composites), new fundamental methods to detect and characterize the probe materials will be developed.

Abstract The objective of this research is to develop ultra-sensitive magnetic resonance imaging (MRI) probes based on MFe2O4 (M=Fe, Co, Mn)-loaded polymer micelles for cancer molecular imaging applications. MRI is a powerful clinical imaging modality that has wide applications in the non-invasive diagnosis and post-therapy assessment for cancer and other diseases. Currently, low sensitivity of detection (approximately 10[-4] M) of conventional MR contrast agents severely limits their applications in monitoring molecular processes in vivo. In this application, we will investigate MFe2O4 (M=Fe, Co, Mn)-loaded polymer micelles as a new multifunctional platform of MRI probes with high sensitivity and biological specificity. Recently, monodisperse MFe2O4 nanocrystals have been successfully developed with fine tuning of particle size below 20 nm. These nanocrystaline materials have demonstrated unique super-paramagnetic (e.g. CoFe2O4) or ferromagnetic (e.g. CoFe2O4) properties. Herein we propose to develop a novel nanocomposite construct consisting of inorganic MFe2O4 nanoparticles loaded inside the hydrophobic cores of amphiphilic polymer micelles. We hypothesize that MFe2O4 -loaded polymer micelles will provide a safe and ultra-sensitive MRI probe for molecular imaging applications. In one specific application, we will functionalize micelle surface with a cyclic Arg-Gly-Asp (cRGD) peptide and examine the imaging of alpha(v)beta(3) receptors on the angiogenic endothelial cells in the tumor neo-vasculature. The specific aims are: (1) Investigate the MR relaxivity and sensitivity of MFe2O4 -loaded polymer micelles; (2) Evaluate the safety and biocompatibility of the MFe2O4 -loaded polymer micelles; and (3) Evaluate the effectiveness of cRGD-functionalized, MFe2O4 -loaded polymer micelles in imaging alpha(v)beta(3) receptors in rabbit VX-2 tumors. This R21 funding will provide critical support to establish MFe2O4 -loaded polymer micelles as a safe and sensitive MR imaging probe for in vivo molecular imaging. If successfully established, these MR probes will provide a powerful technology platform that can exploit the plethora of molecular targets identified in cancer biology for early cancer detection and/or non-invasive assessment of post-therapy outcome.

Award # 3P01DK060564-05S1 Agency NIH Lead Institution Univ of Massachusetts Medical SchoolTypeExtramural

Explanatory Notes This program project combines biochemical, structural and imaging approaches to study proteins by effectively combining diverse experimental techniques with powerful deconvolution algorithms to achieve resolution at the nanometer level.

Abstract Multiprotein complexes that mediate crucial cellular functions, such as signal transduction and membrane traffic, are assembled at the interface of the membrane and the cytosol. The apposition of multiprotein complexes on membrane-cytosol interfaces is achieved in several ways. The best understood manner involves the anchoring of the complex around one or several integral membrane proteins, as is for example the case of complexes of receptor tyrosine kinases and cytosolic signaling proteins. Other multiprotein assemblies are anchored on the polar head groups of phosphoinositides. For example, coat proteins involved in membrane budding assemble onto phospholipid bilayers, and signaling complexes can specifically assemble onto 3' phosphoinositides. The molecular basis for the formation of such phospholipid-based assemblies is poorly understood. This Program Project combines biochemical, structural and imaging approaches to address the question of how multiprotein assemblies are organized around 3' phosphoinositides. The Program Project format is required to effectively combine divergent techniques, which include X-ray crystallography, live cell imaging using digital imaging microscopy with laser-based illumination, and powerful deconvolution algorithms to achieve resolution at the nanometer level. Two model systems will be studied. First, the Czech group will study subcellular localizations of PtdIns(3)P, PtdIns(4,5)P2 and PtdIns(3,4,5)P3 and the multiprotein signaling complexes composed of GRPI, ARF-GTPase, and GRSPI, which are anchored around Ptdins(3,4,5)P3. Second, the Corvera group will study an endosome fusion complex containing EEA1, RabS and calmodulin, which is anchored around PtdIns(3)P. Specific questions to be addressed include: is the interaction of a protein with a phospholipid head group necessary and sufficient to determine its localization to specific regions of the membrane? For this, GFP-fusions of the proteins cited above will be used to determine the subcellular distributions of 3' phosphoinositides in fixed and live cells relative both to the kinases that produce them and to the other components of the complex. As a further means to determine the mechanisms by which these complexes function, the Lambright group will seek to solve the crystal structures of individual protein components as well as of their complexes with lipid head groups. Such structures will be important in defining the spatial topographies of signaling complexes relative to the membranes to which they are bound. The activities in the three projects are highly integrated and collaborative, and are supported by an Imaging Core rated as outstanding in the previous review.

Award # 5R01DK068399-02 Agency NIH Lead Institution Johns Hopkins UniversityTypeExtramural

Explanatory Notes Project supports the synthesis and characterization of nanomaterials targeted for biological study. Nanomaterials are characterized at the tissular, cellular, and subcellular level by various techniques including electron microscopy.

Abstract Liver represents one of the most important targets for gene medicine applications because of the access of the transgene product to systemic circulation, and because it is the site of many metabolic genetic disorders, viral infection and malignancies. A critical barrier in realizing the potential of liver-targeted gene transfer is the development of a safe and efficient gene carrier in combination with a feasible and efficient administration route. To address this issue, we are developing a nanoparticle gene delivery system based on chitosan, which is a biodegradable, biocompatible, and structurally versatile natural polymer. Our recent 'proof-of-concept' experiments have shown that intrabiliary infusion of nanoparticles achieves high levels of transgene expression in rat liver. In this proposal, we build on this finding and hypothesize that high, persistent and liver-targeted transgene expression can be achieved by the systematic optimization of the structure and physicochemical properties of chitosan-DNA nanoparticles and the intrabiliary infusion parameters. The goal of the following set of four specific aims is to experimentally verify this hypothesis and to elucidate its underlying mechanisms: (1). To synthesize and characterize novel chitosan-DNA nanoparticles with optimal complex stability, colloidal stability and hepatocyte-targeting capability. Structure- property-transfection activity relationships will be established; (2). To optimize administration parameters for intrabiliary infusion of chitosan-DNA nanoparticles. Four leading nanoparticle candidates from Aim 1 will be used to evaluate infusion parameters that could significantly affect in vivo transgene expression through intrabiliary infusion; (3). To characterize the nanoparticle/DNA delivery process at the tissue level, at the cellular level in the liver, and at the subcellular level by quantitative PCR, immunofluorescence staining and EM. The information will provide insight to the detailed mechanism of nanoparticle transport and gene delivery through intrabiliary infusion; (4). To validate the gene delivery strategies developed in Aims 1-3 by producing two types of gene product: the systemic circulation of IFN-alpha2b in normal rat, and the correction of a congenital metabolic defect in the Gunn rat mediated by the hUGT1 A1 gene. These experiments will validate our proposed gene delivery strategy and demonstrate its broad applicability for production of proteins intended for systemic distribution and for localized liver-specific diseases.

Award # 5R21MH075059-02 Agency NIH Lead Institution University of California-IrvineTypeExtramural

Explanatory Notes In combination with other intracellular measurement techniques, which may involve nanoparticles like quantum dots, this technique will improve ways of correlating electrophysiological measurements with intracellular biochemical or genetic pathway measurements not currently done.

Abstract The long-term objective of this project is to understand how morphogen gradients pattern the cerebral cortex and other tissues during development. Morphogen gradients are fundamental to animal development, and morphogen defects are primary causes of human birth defects and malformations of the cortex. Nonetheless, tremendous controversy remains about the mechanisms by which morphogen gradients act, which limits our understanding of these human disorders. For the most part, this controversy revolves around a single issue - the inability to distinguish morphogen activities that do not depend on cell-cell communication (the "classical" model) from those that do. To date, insight into this issue has relied on heroic studies using traditional dissociated cell cultures, which are limited both in terms of experimental efficiency and as models of natural morphogen gradients. However, a microfluidic culture device has the potential to address these limitations. This microscale device generates precise and continuous biomolecular gradients with different profiles onto cells, and is designed for time-lapse microscopy. These features should provide several biological and practical advantages over traditional cultures for modeling and studying morphogen gradients. Preliminary studies with cortical precursor cells (CPCs) confirm the promise of this system, but have also identified device design features that need to be optimized in order to answer the basic question driving this proposal - which CPC responses in the normal cortex are determined solely by extracellular morphogen concentration, and which are not? The goals of this R21 proposal are to fabricate optimized microfluidic devices for culturing CPCs (Aim 1) and to develop real time assays with single cell resolution in order to efficiently study CPC responses as functions of morphogen concentration, gradient profile, cell density, and time (Aim 2). To achieve these goals, a multidisciplinary team with primary expertise in bioengineering (Noo Li Jeon, the developer of the original gradient-generating device), morphogen gradients (Arthur Lander) and cortical development (Edwin Monuki) has been assembled. If successful, this proposal should not only advance our understanding of cortical development and malformations, and of morphogen gradients in general, but should also provide a versatile microfluidic tool with a wide range of basic and clinical applications.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes Single-photon spectroscopic techniques are being developed to assess the purity and chemical composition of nano-EHS relevant nanomaterials in complex matrices such as tissues or fluids.

Abstract Multiple spectroscopic techniques are being developed to assess the purity and chemical composition of nano-EHS relevant nanomaterials in complex matrices such as tissues or fluids. These will exploit single or few photon sources and detectors and very small interaction volumes. As a result, the technical challenge is a signal to noise problem, not a diffusion limit problem. This will allow detection of flourescence at extremely low concentrations, which can be significant for biological agents for which on a few cells are at a toxic level.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This project will impact the development of rapid spectroscopic techniques for detecting chemical and biological agents. Such work furthers the ability to detect nanomaterials.

Abstract Knowledge of the photon quantum state is exploited to improve measurement uncertainty from N-1/2 to N-1, thereby increasing the resolution beyond classical limits and/or decreasing the measurement time. This will impact development of rapid spectroscopic techniques for detecting chemical and biological agents.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes New measurement approaches to study novel nanostructured material are a principle component of this project, and support the development of methods to detect nanomaterials.

Abstract We will develop new metrologies and nanostructured materials to enable roomtemperature, high sensitivity thin film magnetic field sensors. These devices will be incorporated into systems for biomedical, homeland security, and magnetic field mapping applications. Room temperature magnetic field sensors are currently limited to relatively high fields (greater than 1 nanotesla). Measurement and mapping of ultra-low magnetic fields (on the order of 1 picotesla) will revolutionize many technologies including DNA sequencing and pathogen detection using sensors for molecular nanomagnet tracers, non-invasive measurement of iron overload in the body, and biohazard detection with sensor arrays for magnetic bead chemical tags.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This project seeks to develop a superresolution optical microscopy platform. This will enable in-situ characterization of organic and biological materials and supports the development of methods to detect nanomaterials.

Abstract The scientific and metrology underpinnings necessary for the realization and operation of superresolution light microscopy in real world applications will be developed and then used to design and build a flexible superresolution optical microscopy platform. When combined with vector point spread function engineering, this platform will establish the state-of-the-art in optical diagnostics for in-situ, characterization of organic and biological materials, and will ultimately represent a field upgrade for conventional microscopy.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This project iinvestigates use of semiconductor quantum nanowires in a variety of sectors. Metrology efforts support the development of methods to detect nanomaterials.

Abstract Semiconductor quantum nanowires (SQNW) offer new revolutionary applications in biological/chemical sensors, in vivo biomedical diagnostics and therapies. SQNW lasers with emission apertures roughly 20-100 nm in diameter (about the size of a virus) could lead to ultra high resolution microscopy and cellular-level imaging. The high surface-area-to-volume ratio, coupled with the electrical sensitivity, may lead to a compact ‘electronic noise’ for chemical and/or biological agents. Besides providing new nanometrology tools for material science and biotechnology, and new sensor capabilities for homeland security needs, our efforts will also result in new nanoscale data and best practices for these areas.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This project is developing new techniques for rapid acquisition of chemical information with high spatial resolution.

Abstract Researchers have developed a method for obtaining three-dimensional broadband vibrational spectra in a microscopic modality. The measured spectra are similar to those obtained from Raman but are acquired with approximately 1,000 times greater efficiency. This new technique is a broadband variant of a well-established nonlinear spectroscopy—Coherent anti-Stokes Raman scattering (CARS). CARS microscopy is becoming increasingly important in biological imaging. Ordinarily, optical microscopy requires researchers to stain (visible or fluorescent) for specific structures or metabolic states. This type of approach cannot be used for all cellular components. CARS microscopy provides a partial solution to this because it is non-invasive and has selective chemical sensitivity; however, its full potential for species-selective imaging usually is limited by restricted spectral bandwidth.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes A wide range of ion, electron, and X-ray spectroscopies, microscopies, and microanalysis techniques are under exploration for the detection and characterization of nanomaterials.

Abstract Cutting-edge electron and ion-based microscopies as well as mass spectrometry techniques will be used for determining nanomaterial shape and structure. Standards for validation of these methods will be developed. Existing techniques for chemical and structural characterization with atomic resolution will be leveraged, and methods that provide information about the chemical structure of individual nanoparticles and the chemical structure of agglomerates of nanoparticles will be developed. Applicable approaches include a range of ion, electron and X-ray spectroscopies, microscopies and microanalysis techniques.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program supports the development of calibration techniques which will allow accurate detection and measurement of nanomaterials.

Abstract For the optics and semiconductor industries, NIST developed a calibrated atomic force microscope, which is calibrated against the wavelength of light in all three coordinate axes. This device calibrates three-dimensional artifacts that, in turn, will be used to calibrate scanning probe microscopes. The semiconductor industry relies on NIST to provide accurate surface measurements of step heights and pitch spacings. For the data storage industry, NIST calibrations support line edge roughness measurements. This program is responsible for certifying Standard Reference Materials for roughness and for calibration of the magnification of scanning electron microscopes.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes Investigations on the possible optical measurements of nanometer-sized features supports the development of methods to detect nanomaterials.

Abstract Optical microscopy will be advanced to unprecedented levels of performance through theoretical and experimental development of a new technique called "scatterfield optical imaging". This new method promises to make possible optical measurements of nanometer-sized features using high-throughput, low cost optical methods with the potential for an enormous impact on innovation and quality control in semiconductor manufacturing and nanotechnology as well as providing the measurement basis for new calibration standards well beyond the state-of-the-art. The new microscopy methods proposed here will enable a significant leap in the sensitivity to geometrical changes of the sample on the order of 1 nm. This method could be developed to image molecular structures.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This project seeks to develop a comprehensive set of metrological tools for measuring nanoparticles in biomaterials and bio-relevant media.

Abstract This program will address critical issues related to the complete cycle of nanoparticles within biosystems. It will accomplish this by developing a comprehensive set of metrological tools for assessing the dimensional and compositional properties of nanoparticles in biomaterials and bio-relevant media. This includes dimensional metrology to obtain precise structural information on nanoparticles combined with image enhancing simulations, and surface chemical characterization methods with atomic resolution.

Award # 0528873 Agency NSF Lead Institution University South CarolinaTypeExtramural

Explanatory Notes The advancement of nano investigative techniques ( including a range of microscopy techniques) are a critical component of this project.

Abstract The project will use nanofabrication methods (such as pulsed laser deposition) and advanced nano investigative techniques (such as SEAM, SPM, TEM, EPMA, EDS, HR-SEM, AFM, XRD) to grow miniaturized thin-film active sensor arrays on typical structural materials for use in advanced structural health monitoring applications. Appropriate buffer interfaces to ensure durable adherence of the ferroelectric film to the structural material and coherent epitaxial growth will be developed. High-performance environmentally friendly ferroelectric compositions will be developed. Modeling and analysis will be used in developing the buffered interface, the nano-fabrication processing methods, and the Lamb/Rayleigh wave phased-array algorithms for structural health monitoring applications of these new devices. The highly interdisciplinary character of the proposal is covered by a sensors small team with members from the disciplines of material science, electrical engineering, mechanical engineering, and physics, spanning over four universities (Louisiana State University, Pennsylvania State University, University of South Carolina, University of Texas at San Antonio), of which one is a Hispanic-serving institution (HSI): The project will employed analytical modeling, numerical simulation, and experimental validation. It will build on the preliminary results obtained by the individual investigators. In additional to technical activities, the project will address important sensors education and outreach activities.

Award # 0547273 Agency NSF Lead Institution Duke UniversityTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract The objective of this research is to enable simultaneous sensing of multiple physical phenomena for application to aerosol detection, homeland security, and medical field-diagnostic tools, for example. The approach is to use hybrid nanomaterials in a monolithic, semiconductor-based heterostructure for multi-functional sensors. Two classes of quantum-confined semiconductor nanomaterials are considered; colloidal quantum dots synthesized by chemical reactions and Stranski-Krastanow quantum dots grown by strained-layer epitaxy. The grand challenges to be addressed are: i) synthesis of colloidal quantum dot/polymer nanocomposite thin-films for optoelectronic device active regions, and ii) synthesis of nanocomposite thin-films featuring disparate nanomaterials embedded in polymers or semiconductors with atomically-sharp interfaces, dopant incorporation capability, and electrical contact layers. A multi-spectral photodiode array synthesized using a hybrid nanomaterial growth system will be demonstrated to culminate the proposed project.

Broader Impacts
The investigation of hybrid nanomaterial device heterostructures for multi-functional sensors with emphasis on environmental applications addresses two 2005 NSF Priority Areas, nanoscale science/engineering and environmental biocomplexity. The proposed hybrid nanomaterial growth system enhances research infrastructure by establishing synthesis techniques for atomically-thin nanocomposite layers. The proposed activities advance discovery while promoting learning by enabling undergraduate research and enhancing the Duke University Electrical and Computer Engineering photonics graduate curriculum. The proposed activities also broaden the participation of underrepresented minorities in science and engineering through the Student Engineers Network, Strengthening Opportunities in Research Saturday Academy and Historically Black College and University student recruitment. Finally, the broad dissemination of results from the proposed activities will be accomplished through scientific publications and the NSF-supported TeachEngineering.com digital library.

Award # 0448796 Agency NSF Lead Institution Boston UniversityTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract The primary objective of this CAREER project is to enable rapid, accurate imaging of thermal and mechanical properties on micrometer and nanometer length scales, thereby contributing to the effort to improve the performance and reliability of micron scale systems and in the effort to understand and control the thermal and mechanical behavior of materials at the nanoscale. The PI develops novel photoacoustic and photothermal (PA/PT) measurement capabilities for the noncontact and nondestructive imaging of surface properties and subsurface defects at the nanoscale, along with an understanding of how the measured quantities relate to the thermo-mechanical properties of the target material. A technique for producing highly localized heat sources and probes for PA/PT imaging is explored, with the goal of pushing the lateral resolution of PA/PT microscopy to the nanoscale regime. This novel imaging modality makes use of the high absorption and local field enhancement around nanoparticles excited at the plasmon resonance frequency, and will have a broad impact in the quest to understand and characterize material behavior at the nanoscale. Next, A high-resolution photoacoustic microscopy system is developed which uses a high-frequency (GHz) modulated excitation source. Improvements in signal to noise ratio over pulsed systems are expected through bandwidth reduction and pulse coding techniques, expanding the range of applications for which this NDE technique is suitable. The final thrust area in this proposal focuses on the modeling of laser generation of acoustic and thermal waves in complex materials systems, an area which is essential for the interpretation of thermal and acoustic signals in these systems, and for accurate inversion of these signals to obtain material property information. The research results will have direct applications in nanofabrication: for online monitoring of the fabrication process as well as in the measurement of the fundamental properties of "as-fabricated" materials systems. This project supports both graduate and undergraduate researchers, and trains these students in a wide cross-section of engineering science that this project draws upon including NDE, applied optics, elastic wave propagation, and nanoscale engineering.

This CAREER project supports the development and implementation of the Learning Experiences for New Scientists (LENS) program in which academically at-risk 7th and 8th grade students come to Boston University for a week of hands on demonstrations and problem solving activities. This program targets students at a critical time in the development of science skills, and is aimed at reducing the number of students graduating from high school who lack basic proficiency in science and math. The multi-media presentations developed for the LENS program are placed on the Web to disseminate to other educators, and are made available for other outreach programs through the Learning Resource Network (LERNet) program at Boston University. The PI has arranged for a graduate student to work at Lawrence Livermore National Lab for one summer under this project. This opportunity allows the student to interact with several experts on laser ultrasonics at LLNL, and to broaden his/her educational background. The PI develops a new undergraduate course in the area of mechanical behavior of materials, in which the NDE of materials will be discussed. The PI also develops NDE demonstrations for a graduate course in experimental techniques in solid mechanics. The PI serves as the faculty advisor for a professional society student group, and will host a future regional student conference at BU. The PI plans to invite representatives from local companies and educational institutions to speak to the students about the importance of NDE

Award # 0552772 Agency NSF Lead Institution Wayne State UniversityTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract This renewal award provides support for a three-year REU Site at Wayne State University. The program builds upon the current education and research theme of the previous REU program on Smart Sensors and Integrated Devices (SSID), and also extends it to a systems approach of nano-integration with micro-systems research. The program will provide 10 students each year with research experience in cellular biophysics, biosensing science and technology, nanoscale devices, and translational applications in medicine and environmental sciences. REU participants will work closely with an interdisciplinary team of students with at least two faculty advisors to understand, develop and apply new sensor technologies starting from one or more sensing mechanisms (optical, mechanical, electrical/electromechanical, and magnetic) and extending to nano integration and biofunctionality of the sensor/biological interface.

The main objectives of the program are to provide undergraduate students with a comprehensive research experience in Nanoscale Structures and Integrated Biosensors (NSIB) and make them aware of the opportunities in graduate programs in science and engineering, or in industrial research settings. The program will place emphasis on the participation of female and underrepresented minority students, and encourage the students to pursue graduate programs in physics and engineering.

Award # 0522656 Agency NSF Lead Institution Rensselaer Polytechnic InstituteTypeExtramural

Explanatory Notes This work is aimed at developing a highly efficient chromatographic chip system able to separate nanoparticles, detect the presence of nanoparticles, and prepare samples by particle size for further chemical and biological characterization.

Abstract This project will employ microfabrication, selective filling and sol-gel nanotechnology to develop a variety of highly efficient chromatographic chip systems for bioseparations applications. Preliminary results indicate that a novel selective filling technique can be employed to create complex patterns in chromatographic chip systems. State of the art techniques employed for making sol-gel binders with unique properties will be developed for chromatographic chip systems. Efficient chromatographic chip systems will be developed by the immobilization of small particle diameter chromatographic materials using sol-gel techniques along with increased column lengths and gradient operation. The chromatographic chip systems will be evaluated for pore morphology, permeability, pore size distribution, chromatographic efficiency, and separation capability using biological test mixtures. Multifunctional systems will be examined using the selective filling sol-gel technique for integrating enzymatic digestion with gradient chromatography in a chip format. In terms of the broader impacts, this project may guide the development of a new platform technology for implementing chromatographic separations in multifunctional chip formats, which will have a potential impact on a variety of fields ranging from proteomics to environmental science. In addition, the development of selective filling sol-gel immobilization technology will facilitate advances in the state of the art of lab on chip devices. The project will also educate both undergraduate and graduate students in the state of the art of sol-gel nanotechnology as well as chip based separation systems. Chromatographic chip technology can impact areas where bioseparations are important, including environmental science and proteomics

Award # 0425780 Agency NSF Lead Institution University of PennsylvaniaTypeExtramural

Explanatory Notes This work is targeted at developing new instrumentation for detection and characterization of nanoparticles in biosystems.

Abstract The University of Pennsylvania's Nano Science and Engineering Center on Molecular Function at the Nano/Bio Interface will exploit Penn's strengths in design of molecular functionality, quantifying behavior of individual molecules, and interactions at organic/inorganic interfaces to perform research that establishes the foundation for understanding molecular function in the context of interfacing with physical systems. The NSEC unites 18 investigators from three schools (the School of Engineering and Applied Science, the School of Medicine, and the School of Arts and Sciences). Two multidisciplinary research teams are focused on aspects of the fundamental issues outlined above. Additionally, two cross cutting initiatives develop ideas integral to the research themes and make explicit links between them. The two fundamental themes are: optoelectronic function in synthetic biomolecules and mechanical motion of molecules from physiological systems. The two cross-cutting initiatives are on Molecular Nano Property Probes and Ethics in Nanotechnology.

The impact of these efforts will be felt in biophysics, bioengineering, chemistry, electrical and mechanical engineering and materials science. Discoveries from this effort will provide a sound basis for the development of new technologies for nanoscale device manufacture, drug delivery and integrated chemical sensors, enabling several near term practical applications as well provide the basis for future practical implementation. Furthermore, these issues are also at the core of understanding many complex biological/physiological processes.

The broad impact of the NSEC will occur on several levels. From a technical perspective it will articulate the critical issues that define the field at the interface of nanotechnology and biology at the molecular level. As such it will focus the attention of many disciplines to an area that is at the core of future of the field. The NSEC will impact public education, social discourse, workforce development and diversity, both locally and nationally. The implementation of educational activities in an urban environment will target a highly diverse audience at the early stage when exposure to exciting science can influence interests and future career choices, while developing models that can be implemented across the country. This NSEC will take a leadership position in the social discussion of ethics in nanoscience and technology

Award # 0084173 Agency NSF Lead Institution Florida State UniversityTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract The National High Magnetic Field Laboratory (NHMFL) is operated by a consortium composed of the University of Florida, Florida State University, and Los Alamos National Laboratory. Florida State University administers the Laboratory as a national user facility, available competitively to users on the basis of merit. Established in 1990 and dedicated in 1994, the Laboratory through an extraordinary state-federal partnership with multi-agency participation has developed unique facilities in support of magnet-related research at the highest attainable magnetic fields. The Laboratory is structured around four major thrusts: (1) user facilities
developed in response to users' needs that are opening new frontiers for science opportunities, (2) magnet science and technology developed in partnership with the private sector to enhance U.S. competitiveness, (3) basic science research driven by a partnership between external and in-house users that drives new opportunities in high magnetic field science and technology, and (4) the integration of research and education at all levels, and partnership with academia, industry, government and international institutions to advance research and technology in the area of high magnetic fields.

Over the first ten years the NHMFL has put in place a unique range of instruments and facilities for research in high magnetic fields, including continuous field, pulsed fields, and magnetic resonance research. During this time the Laboratory has established itself as the world's leading center for multi-disciplinary research using high magnetic fields. It has developed an outstanding educational program and built strong collaborations with academic, industrial, government and international partners. The focus of the Laboratory is now shifting from a primary emphasis on magnet technology and development to include increased support for service to users from a wide range of scientific and engineering disciplines. The Laboratory is now building on the federal and state investment to realize the full scientific potential of the new facility, while the science and magnet technology programs expand the current capabilities, develop new magnet systems, and drive new science discoveries at the highest fields and at extremes of pressure and temperature. The Laboratory provides continuous fields (up to 45 tesla) in the magnetic field region formerly thought to be reserved for only pulsed magnets, and reversible pulsed fields (60 tesla for tenths of a second and up to 79 tesla for milliseconds). The availability of opportunities in magnetic resonance at 900 MHz and beyond will be a critical aspect of the Laboratory's efforts to build a users' center of excellence in very high field magnetic resonance spectroscopies. The in-house science program has been developed in cooperation with the external users and addresses a wide range of research areas including highly-correlated electron systems, magnetic materials, magnetic resonance spectroscopies applied to the chemical, physical, and biological sciences, and the development of novel instrumentation to take advantage of the magnet facilities available to users

Award # 0114372 Agency NSF Lead Institution University of Minnesota-Twin CitiesTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract This IGERT program focuses on nanoparticle science and engineering, an inherently highly interdisciplinary field that requires researchers with a broad knowledge base of both fundamental scientific and engineering issues. The program addresses the lack of a coherent and well-organized Ph.D. training in this field. Faculty from five departments and six graduate programs at the University of Minnesota have come together to develop an interdisciplinary program transcending departmental boundaries to meet this challenge. At the core of the educational approach is the establishment of a new graduate degree program - a freestanding graduate minor program in Nanoparticle Science and Engineering. Several new interdisciplinary core courses will be developed to offer students a coherent and comprehensive set of courses. Students enrolled in the IGERT program will participate in interdisciplinary research training in research groups that include faculty and students from various departments. Nanoparticle research will be conducted in five focus areas: Two areas will address the development of enabling computational and characterization tools. These areas will form the foundation for research in application oriented areas focusing on new materials, devices, and the environment. Exceptional career development opportunities will add to the appeal of the IGERT program. These include an internship program in corporate and government laboratories, international exchange opportunities, attendance of scientific meetings at an early stage in the career, an annual interdisciplinary symposium, leadership and technology management courses, and training in ethical conduct of research. IGERT Fellows will use the excellent infrastructure at the University of Minnesota, such as its Supercomputing Institute, its Microtechnology Laboratory, and its Characterization Facility. A unique collaboration with Florida A&M University focusing on the preparation of minority undergraduate students for graduate studies and their recruitment into the IGERT program will enable building of a diverse student body.

Award # 0522005 Agency NSF Lead Institution Wayne State UniversityTypeExtramural

Explanatory Notes Efforts targeted to investigation and design of nanostructured molecular sieves using molecular and atomistic simulation for detection of toxic industrial materials at the nanoscale or molecular level.

Abstract Semi-conducting metal oxide (SMO) based sensors for the detection of chemical warfare agents (CWA) and toxic industrial materials (TIM) exhibit sensitivities on the order of parts per billion. SMO based sensors have potential advantages over other methods of chemical agent detection in terms of size, weight and cost. One limiting factor in the use of SMO based sensors for CWA/TIM detection is that these materials are non-selective. That is, there are many molecules, in addition to the agent of interest, which will yield positive detection results. The proposed work is focused on improving the selectivity of semi-conducting metal oxide sensors and the reduction of false positive responses.

Controlling the adsorption of CWA/IM onto the surfaces and their subsequent diffusion through the pores and key factors in the design of such devices. This can be done via a pre-filtering scheme, where a mixed gas stream in passed through a ceramic membrane or activated carbon. Pore size, shape and chemical composition can be tailored to selectively adsorb the molecule of interest. After being concentrated in the pre-filter, the CWA/TIM is released by a chemical displacer or a thermal pulse and sent to the SMO for detection. An alternative approach to pre-filtering is to use templating to form porous semi-conducting metal oxides with high selectivity to the target molecule. In the proposed work, molecular simulation is used to determine the absorption behavior of CWA and their simulants in the molecular sieve MCM-41.

The difficult nature of performing experiments on CWA/TIM motivates the proposed use of computational methods. Molecular simulations is well suited to the study to the toxic material and can be used to extract information on the roles specific intermolecular interactions play in the adsorption process. Because appropriate models (force fields) so not exist for the molecules of interest, significant effort is proposed on the development of transferable united-atom force fields for organophosphates, including the chemical warfare agents sarin and VX. The development of molecular models is a required first step in use of simulation for the design of SMO based sensors. These molecular models will allow for the use of simulation to investigate the effects of pore size, shape and composition on the selectivity of porous materials with respect to specific CWA/TIM. Atomistic simulations and ab initio methods are used to identify specific porous structures (shape/size) with the high selectivity necessary for the sensing of CWA/TIM with low false positives.

Broader Impacts

Recent world events, such as the release of sarin gas into the Tokyo subway system and the current concern over potential adversaries suspected development and use of chemical and biological warfare agents underscore the importance of developing highly mobile, accurate sensors and decontamination equipment for these materials. As an outcome of the proposed research, the PI expects to overcome the current limitations of the field by developing the necessary computational infrastructure for the use of simulation in the design of novel templated molecular recognition materials. These templated molecular recognition materials hold the promise of high selectivity and sensitivity to chemical warfare agents compared to other porous materials. Development of adsorbents with high affinity to a specific target molecule is expected to result I improved sensors, filters and catalytic materials. Such developments are expected to reduce the threat of the use of chemical warfare agents by terrorist organizations, improving national security and public health. Furthermore, accurate force fields describing the interactions between quest molecules and metal oxide surfaces will allow other research groups to use molecular simulation as a too to design novel adsorbent and catalytic materials for other purposes

Award # 0531171 Agency NSF Lead Institution University of Massachusetts-AmherstTypeExtramural

Explanatory Notes Metrology efforts support the development or improvement of methods to detect nanomaterials.

Abstract This Nanoscale Science and Engineering Center (NSEC) is a comprehensive research and education platform that will stimulate U.S. competitiveness by moving nanotechnology from laboratory innovation to manufacturable nanostructured components and devices. The Center for Hierarchical Manufacturing builds on recognized excellence in nanoscience and technology at UMass and a world-class program in polymer research to yield complete specification of nanostructures combining directed self-assembly and imprinting with subsequent transfer of 2-D and 3-D structures, and advanced deposition techniques, into active components, functional materials, and fully-integrated devices. Bottom-up processes will be seamlessly integrated with conventional fabrication methods for dramatic advances in semiconductor devices, microelectronics, biomedical applications, and other areas. In addition, the Center offers a new strategic model to bridge the innovation-to-implementation gap through test beds that combine leading breakthrough technology, professional market analysis, industrial-scale fabrication processes, and facilitated technology transfer.

The impact of this Center is strengthened through strong collaborators including leading R&D consultant TIAX and prototyping partner Lucent Technologies will allow the Center to drive concepts to commercialization. Students educated in this environment will be well prepared for careers that partner innovation with implementation. The Center initiates a National Nanomanufacturing Network (NNN)--a catalyst for U.S. nanomanufacturing-based economic development, a network of shared manufacturing facilities, a dynamic web-based information clearinghouse, and a pathway for university-industry-government partnerships. The NNN will integrate and amplify the impact of all NSECs and the nanomanufacturing research community. The Center will address a national need by creating and disseminating research-based multimedia instructional materials to stimulate and educate audiences ranging from K-12 students and teachers, community college learners, and the public. A strong societal implications program includes national workshops and survey studies. The Center's efforts are leveraged by strong commitments from the University of Massachusetts Amherst and the Commonwealth of Massachusetts.

Award # 0118025 Agency NSF Lead Institution Northwestern UniversityTypeExtramural

Explanatory Notes These efforts aim to develop state-of-the-art instrumentation for detection of nanoparticles with high accuracy and sensitivity.

Abstract This award establishes a Nanoscience and Engineering Center (NSEC) for Integrated Nanopatterning and Detection Technologiesat the Northwestern University. The Center will develop state-of-the-art nanopattering capabilities that are compatible with soft materials and can be used for the development of powerful new detection systems. Several emerging patterning tools and synthetic methods will be developed to fabricate nanostructured materials and devices. The Center's research strategy focuses on (1) the science and technology of surface template-driven assemblies that rely on chemical and biochemical recognition events, and (b) the experimental and theoretical study of signal transduction in soft matter nanostructures. The research program is organized under three themes: (1) Chemical and Biological recognition; (2) Nanostructured Assembly; (3) Transport and Transduction in Nanostructures Formed via Surface-Directed Assembly. As the research progresses, focus will be placed upon the development of biological (nuicleic acids and proteins) and chemical (small molecule) sensors. The research outreach will include University of Chicago, University of Illinois, Urbana-Champaign, Harold Washington University, and Argonne National Laboratory. The Center will establish formal programs with international comp[anies, universities, and laboratories in the area of nanoscale research. The Center will take a lead to establish the first global network of centers and institutes in the area of nanoscience and engineering. In the area of undergraduate education, the Center will establish Research Experience for Undergraduates (REU) and the Minority Internships in Nanotechnology (MIN) programs. The MIN program will be promoted through the minority institutions in the area. Middle and high school teachers will be provided research experience through Research Experience for Teachers (RET) program. A web-based highly flexible "Functional Nanostructure" module will be developed and distributed. New courses in nanotechnology will be developed for undergraduate and graduate levels. Industry participates in the Center activities by providing company personnel as student mentors, and by supplying source codes for software development. The Center has a program to advice on formation of startup companies.

Award # 0448835 Agency NSF Lead Institution California Institute of TechnologyTypeExtramural

Explanatory Notes This work is targeted toward understanding and developing new principles and toward the construction of versatile and inexpensive biosensors with nanoscale sensitivity.

Abstract The Pierce Lab at Caltech proposes a research program that combines theoretical, computational
and experimental components to advance the field of structural nucleic acid nanotechnology in developing rigorous and robust methods for encoding arbitrary mechanical function in nucleic acid sequences. Modeling and algorithm development projects will address key analysis and design challenges. The investigation of pseudoknot biophysics will help to create a physically based potential for partition function algorithms and illuminate the role of topology in the behavior of the "kissing hairpin" mechanism under experimental study for catalytic fuel delivery to autonomous devices as well as:
-on implementing an efficient hierarchical mutation algorithm for optimizing affinity and specificity for target secondary structures using partition function information
-on methods for identifying and designing multiple features of an energy landscape including networks of metastable states relevant to autonomous device design
-and on formulating and optimizing the multi-objective design problem that characterizes device engineering, in which multiple strands must conditionally adopt different structures depending on the subsets of strand species that are present.

Intellectual Merit
The proposed research program seeks to develop analysis and design tools that will dramatically increase the level of rigor, accuracy, and automation with which nucleic acid devices are engineered. The work encompasses physical modeling, algorithm development, mechanism design, and experimental validation, with the unifying goal of enabling the manipulation of subtle features in the multiple energy landscapes that underlie the design of devices with many conditionally stable states. The development of a practical catalytic fuel delivery method and subsequent construction of a robust autonomous walker that can take hundreds of unassisted steps would represent a breakthrough in the field. Likewise, HCR provides a new functional paradigm for nucleic acid nanotechnology, creating many attractive avenues for investigation.

Broader Impact
The ability to engineer autonomous devices at the nanometer length scale may someday have importantmedical applications including the targeted delivery of toxic drugs. In the nearer term, HCR has the potential to facilitate the construction of versatile and inexpensive biosensors. Undergraduates have the opportunity to participate fully in this interdisciplinary research through the Caltech Summer Undergraduate Research Fellowship program. Researchers at other universities can freely download source code for the analysis and design software developed in the Pierce Lab. At Pasadena public schools, 62% of students participate in free or reduced-price meal programs and more than 80% of students are underrepresented minorities. For many students, the Jr. Teacher program will provide a rare opportunity to consider the attractions of pursuing a career in science or engineering.

Award # 0425914 Agency NSF Lead Institution University of California-BerkeleyTypeExtramural

Explanatory Notes Projects are targeted to the development and use of two closely related nanosensor systems: (1) a new personal and community-based sensor for the environmental monitoring of nanoparticles, and (2) technology for the chemical/biological sensing of nanoparticles with integrated communication and power for tagging, tracking, and locating applications.

Abstract The mission of COINS is to inspire and realize truly revolutionary applications involving molecular transport, replication, and energy conversion using nano-mechanical technology, integrated with suitable societal implications studies and educational, outreach, and knowledge transfer programs. Specifically, the technical focus of COINS is to develop, in parallel, two closely related nanosensor systems: (1) a new Personal And Community-based environmental MONitoring (PACMON), and (2) a chemical/biological sensing with integrated communication and power for tagging, tracking, and locating applications (TTL).
To realize efficiently the goals of COINS, a carefully designed and integrated plan combining the talents of 30 investigators spanning 10 different disciplines (applied physics, bioengineering, chemical engineering, chemistry, economics, electrical engineering and computer science, materials science and engineering, mechanical engineering, molecular and cell biology, and physics) has been established and set into motion. Six major research thrusts - synthesis, simulation, characterization, instrumentation, integration, and society - couple the three naturally divided technology tiers of System Integration, Enabling Technology, and Fundamental Knowledge Base, all necessary to support TTL and PACMON.

Initial abstract (in 2004)
The Nanoscale Science and Engineering Center entitled Center for Affordable Nanoengineering of Polymer Biomedical Devices (CANBD) is a partnership between the U. of Akron, Boston University, UC Berkeley, Johns Hopkins, Florida A&M, and Purdue. The NSEC includes 38 investigators from 9 departments.

The Center seeks to develop polymer-based low-cost nanoengineering technology that can be used to produce nanofluidic devices and multifunctional polymer-nanoparticle-biomolecule nanostructures for the next generation medical diagnostic and therapeutic applications. The research plan is comprised of three thrust areas. The Nanomanufacturing Thrust Area combines 'top-down' fabrication and 'bottom-up' molecular self-assembly to produce well-defined passive and active nanostructures. In the Transport Phenomena Thrust Area, the research will achieve design capabilities at the nanoscale by combining nanofluidic design, transport phenomena at the nanoscale, and multiphase transport structures with multiscale modeling and macroscalar property assessment. Biocompatibility issues will be addressed in the Biocompatibility Thrust Area in parallel with the development of new nanofluidic designs and devices.

The near-term goal of the three closely linked research thrust areas is to design and fabricate polymer-based, 3D nanofluidic circuits for manipulating the shape, orientation and transport behavior of individual biomolecules in well-defined nanoscale flow fields (5-100 nm). Test bed examples include a simple, handheld protein separation/diagnostic device; a nanoneedle cell patch for low-invasive delivery of genes and macromolecular medicines into cell walls; and biomolecular nanopumps as synthetic ion channels. The ultimate goal is to design and assemble a nanofactory based on the integration of nanofluidic circuits, synthetic chemistry and biological complexation.

Center collaborators include at least 20 companies in Ohio and the U.S., Battelle, the Cleveland Clinic Foundation, the National Cancer Institute, Oak Ridge National Laboratory, Wright Patterson Air Force Labs, and researchers in Asia, Australia and Europe. The Center also plans to coordinate closely with NSECs at the University of California at Los Angeles and the University of Illinois-Urbana (nanomanufacturing), the NSF STC at the University of North Carolina at Chapel Hill (environmentally responsible solvents), and the NSF ERCs at the University of Washington (biomaterials and biocompatibility) and the Georgia Institute of Technology (3D tissue models) because of complementary research agendas.

The education and outreach vision of the Center is to integrate the latest research developments into a practical student curriculum that imparts multidisciplinary skills and global awareness to both graduate and undergraduate students. The key education elements include a series of new courses to introduce nanoengineering of biomedical devices and related topics; an interdisciplinary curriculum offering an undergraduate minor and a graduate certificate; internships and visits to industry and national laboratories in the U.S. and abroad; and web-based dissemination. The recruitment and retention of minorities and women will be emphasized through close collaboration with minority institutes such as FAMU/FSU. Undergraduate students will participate in research via senior honors theses and targeted REU support. Outreach activities include web-based science modules for K-12 students nationwide; workshops and short courses for high school science teachers and industrial researchers; and on-site research projects and workshops for middle school and high school students supervised by graduate students.

Award # Agency DOE Lead Institution University of RochesterTypeExtramural

Explanatory Notes This project seeks to understand nanoscale material optical phenomena through the detection of a single molecule in a complex environment. This work provides a basis for the development of novel schemes and techniques for molecular identification and analysis, and supports efforts to understand nanomaterial structures, properties and behavior.

Abstract Our studies will concentrate on the interaction with single molecules because they are well defined systems in terms of their chemical structure and electronic/vibrational states. Although there are many unresolved questions related to their statistical behavior at ambient conditions, single molecules are quantum systems that are well approximated by elementary two-level systems. As such, they are ideal probes for measuring optical field distributions, and for investigating lifetime variations and emission properties near nanostructures. Once we understand the interaction with single molecules we are in a better position to understand interactions with more complex systems such as molecular aggregates, proteins in biological membranes, or semiconductor nanostructures. We will continue our collaboration with different DOE laboratories to make use of state-of-the-art nanofabrication instrumentation and ultrafast time-resolved measurements.
Our method is well suited to investigate optical interactions that are mediated by virtual photons (van der Waals/Casimir forces), to study electromagnetic friction due to locally fluctuating fields, and to understand the origin of surface enhanced Raman scattering (SERS). Furthermore, the strong field gradients near a metal tip are promising for spectroscopic measurements beyond the standard dipole selection rules. We will use the results from our ongoing DOE project to localize the light-molecule interaction by ’engineering’ the physical properties of a metal tip. We will study the strength of the local field enhancement and investigate the molecule’s radiative decay rate as a function of tip proximity and orientation of the molecular transition dipole moment. The ultimate goal is to develop the ability to detect a single molecule in a complex environment with a spatial resolution in the order of 10-20 nm. The acquired understanding of this project will provide a basis for the development of novel schemes and techniques for molecular identification and analysis.

Award # Agency DOE Lead Institution Pacific Northwest National LaboratoryTypeExtramural

Explanatory Notes This is fundamental research directed toward understanding and controlling the chemical properties of nanomaterials. The goal is to understand the mechanism responsible for the overall particle reactivity and reaction selectivity of reactive metal and oxide nanoparticles. This supports efforts to further understand nanomaterial structures, behaviors, and modifications.

Abstract This research proposal seeks to continue fundamental research directed toward understanding and controlling the chemical properties of reactive metal nanoparticles. The program integrates: 1) synthesis of well defined particles, 2) characterization of their surface and bulk composition as well as physical and electronic structure, 3) measurements of reactivity in vacuum and solution, and 4) theory and modeling work the helps identify reaction mechanisms and charge transport properties of oxide structures. Research from several directions indicates that the shells or coatings on the particles play a major role in determining reaction pathways. Therefore a major goal of this proposal is obtaining an understanding how shell properties control the reaction process and to use that understanding to synthesize particles with optimized properties.

Award # Agency DOE Lead Institution Lawrence Livermore National LaboratoryTypeExtramural

Explanatory Notes This program supports a quantitative investigation of the effects of different parameters on the properties of nanoparticles and enables a better understanding of the behavior of nanomaterials as a function of their physical properties.

Abstract The focus of this research is the quantitative investigation of the effects of size, surface composition, proximity, and dopants on semiconductor nanoparticle properties. We employ unique methods of particle synthesis that allow the greatest control over these physical parameters and utilize state-of-the-art synchrotron radiation-based characterization methods that provide element-specific atomic and electronic structure information. This powerful combination has proven most valuable to the condensed matter theory community who have compared the experimental data to their computation results with and, as a direct result, helped refine our understanding of quantum dot structure/property relationships and guided new material discovery.

Award # Agency DOE Lead Institution University of HoustonTypeExtramural

Explanatory Notes This project supports work on the the determination of the structure of nanophase clusters. Metrology efforts under invistigation provide developments or improvements to methods to understand the interaction of modifications with properties of nanomaterials, a focus for this research need.

Abstract This project is a continuation of our work on the structure of nanophase clusters in zeolites and two additional topics. We will conduct anomalous X-ray scattering studies on Hg-Se in the cages of Nd-Y followed by Hg-Se but in a LTL zeolite with one-dimensional tubular channel. The comparison of this semiconductor nanocluster in these two distinct environments should prove fascinating especially as they are of appreciable technical interest. In addition, Raman scattering will be done with Dr. Milko Iliev of the Texas Center for Superconductivity and Advanced Materials in order to correlate the molecular structure with the electronic structure of these caged semiconductors. In addition to the zeolite work we will 1) conduct a detailed study of amorphous CaCO3 (ACC) which turns out to serve as the precursor to a rich variety of biological and geological materials and 2) with Dr. David L. Price we plan to do containerless levitation of Fe-AL and Ti-Al liquid metal alloys to obtain accurate S(Q)’s including the under-cooled state. Small angle X-ray and neutron scattering (SAXS and SANS) will also be done to explore the issue of defect-induced density fluctuations. This work will include Dr. J. L. Robertson.

Award # Agency DOE Lead Institution University of Tennessee TypeExtramural

Explanatory Notes This project supports development and deployment of new methods and instrumentation to study nanomaterials and nanostructures. The advanced tools may futher the ability to measure properities of complex materials in relation to structures of other features.

Abstract This project supports scientists from the University of Tennessee (T.A. Callcott), and Tulane University (D.L. Ederer) to install and commission a new Soft X-ray Spectroscopy endstation at the Advanced Photon Source (APS) at Argonne National Laboratory and to use this instrumentation to undertake initial studies on complex magnetic materials and nanostructures. The facilities will be used to study the electronic states, including orbital, charge and spin ordering, of complex magnetic materials and magnetic nanostructures. These materials present both great scientific challenges and exceptional technological opportunities. Their characterization and theoretical description is at the frontier of much of present condensed matter physics, while increased understanding opens the prospect of “engineering” these materials for practical applications as sensors, actuators and spintronic devices. The methods of soft x-ray emission and absorption spectroscopy have unique characteristics that enhance their value for the study of complex materials. The new endstation, combined with the soft x-ray circularly polarized undulator (CPU) of the APS, will provide the world’s best facility for the study of complex magnetic materials using SX spectroscopy. A major focus of our proposal is the use of these recently developed techniques and new equipment to fully exploit the ability to measure orbital and spin ordering and correlation effects. Finally, we propose a closely coordinated theoretical effort, which provides essential support for applying the unique capabilities of SX spectroscopies to the challenging problems of complex materials.

Award # Agency DOE Lead Institution University of VirginiaTypeExtramural

Explanatory Notes This project seeks to provide instrumentation to routinely determine the mechanical (and physical) properties of nanoscale materials. Application of the technique to a number of model experimental systems and situations will further efforts to better understand the effects of modifications on the physical properties of nanomaterials.

Abstract It is well known that one of the primary inelastic scattering processes that occurs as fast electrons travel through a thin specimen in the transmission electron microscope (TEM), is the creation of quantized collective electron excitations, know as volume plasmons, in the specimen. Volume plasmon peaks occur in the low-loss region of electron energy-loss spectra (EELS) from all materials and this makes them potentially attractive for determining various material properties, particularly since they are associated with the bonding electrons that are responsible for most material properties. Recent results from our laboratory and in the literature indicate that it is possible to determine a number of important mechanical properties of nanoscale materials, such as their bulk, shear and elastic moduli from their volume plasmon energies using scaling relationships. It is quite difficult, if not impossible, to obtain such properties for nanoscale structures by other techniques, making nanoscale materials characterization using plasmon energies in EELS highly attractive. Nanoscale structures are also particularly well suited for analysis in a field-emission gun TEM, where subnanometer electron probes are readily obtained. However, a number of questions need to be answered in order to utilize plasmon peaks to routinely determine the mechanical (and physical) properties of nanoscale materials, and that is the purpose of this research. The main objectives of this research are to: 1) understand the relationship between the volume plasmon peaks in EELS and the mechanical (and physical) properties of materials on a fundamental level, 2) establish the relationship between the mechanical properties of nanoscale materials and their plasmon energy as a function of material size, so that plasmon measurements in EELS can be used to routinely determine the mechanical properties of nanoscale materials, and 3) apply the technique to a number of model experimental systems and situations in order to understand the accuracy and limitations of the technique. One of the major theoretical accomplishments of this project so far, has been to demonstrate that the strong scaling correlations that exist between the volume plasmon energy obtained experimentally by EELS, and material properties such as the cohesive energy, valence electron density, elastic constants and hardness of various materials, are a direct consequence of the universal binding energy relation (UBER). This fundamental relationship provides the theoretical basis necessary to apply EELS and energy-filtered transmission electron microscope (EFTEM) imaging to determine multiple solid-state parameters of materials at the nanoscale. As a major experimental accomplishment, we have produced the first plasmon-ratio maps of the bulk modulus, cohesive energy and bonding electron density in a material, with sub-nanometer spatial resolution. This accomplishment is significant, because it demonstrates that EFTEM can be used for quantitative material property determination and imaging, establishing a new capability in electron microscopy and materials research. Reference: J. M. Howe and V. P. Oleshko, "Application of Valence Electron Energy-Loss Spectroscopy and Plasmon Energy Mapping for Determining Material Properties at the Nanoscale", J. Electron Micros., vol. 53, 339 (2004).

Award # Agency DOE Lead Institution Washington State UniversityTypeExtramural

Explanatory Notes This project supports the examination of nano-structures and their properties using novel instrumentation, which may ultimately provide new avenues to investigate how modifications affect nanomaterial properties.

Abstract Monoenergetic positrons from beta decaying radioactive sources carry a net polarization proportional to the initial velocity of the positrons which is maintained through the moderation process (to generate a monoenergetic beam) and when they are implanted into a sample. Given an asymmetry in the spin density distribution where the positron eventually annihilates, this can be observed in the amount of three photon annihilations vs. two photon annihilations. In the absence of positronium (3/4 of which would annihilate into 3 photons in vacuum) the three photon fraction is small but measurable. We have devised two methods to observe small 3 photon components among a vast number of 2 photon annihilations with a sensitivity of about 1 in 400. Using these methods we expect to observe magnetic properties of nano-scale structures such as magnetic NdFe melt spun nano-clusters. To check the sensitivity of the three photon detection apparatus has been constructed and we have measured the ratio of 3 to 2 photon annihilations in a range of samples (metals, semiconductors, insulators and porous materials). Small variations but significant differences have been found. More work is under way to eliminate possible systematic effects as the result is contrary to predictions made for the last 50 years.

Award # 3P42ES004705-19S10031 Agency NIH Lead Institution University of California-BerkeleyTypeExtramural

Explanatory Notes This project provides insight into the processes occurring on or near ZVI surfaces by using techniques designed to probe the surface, such as potentiometry, surface-enhanced Raman spectroscopy and electrochemical quartz microbalance methods.

Abstract The main objective of the proposed research is to assess the potential for using oxidants produced during the corrosion of granular and nanoparticulate zero-valent iron (ZVI) by oxygen to remediate contaminated groundwater and soil. This objective will be realized by studying the reaction mechanisms involved in oxidant production and contaminant transformation and the efficiency of potential treatment methods under conditions similar to those that are likely to be employed in treatment systems. The overall hypothesis that we aim to test is that the oxidative ZVI system offers a practical, cost-effective means of remediating contaminants that have the greatest impact on human health at Superfund sites. Our investigation of the reaction mechanisms will focus on the role of solution chemistry and surface structure on the rate of contaminant transformation. To gain insight into the processes occurring on or near ZVI surfaces, chemical processes occurring in the solution phase will be measured in conjunction with studies conducted using techniques designed to probe the surface, such as potentiometry, surfaceenhanced Raman spectroscopy and electrochemical quartz microbalance methods. Our investigation of the potential applications of the oxidative ZVI system to contaminant remediation will focus on permeable reactive barriers and water infiltration systems used to treat organic contaminants and drinking water treatment systems used to remove arsenic. These studies will extend the research in oxidant formation mechanisms to account for the effect of oxide coatings on the ZVI surfaces on contaminant oxidation rates and transport of contaminants to and from the corroding iron surfaces. This research has the potential to provide innovative and cost-effective ways of removing contaminants from groundwater and drinking water that are difficult or expensive to treat by conventional methods. The development of these technologies could reduce human exposure to organic and inorganic contaminants of concern

Award # 5R21DK072450-02 Agency NIH Lead Institution Northwestern UniversityTypeExtramural

Explanatory Notes This project studies the effects of modifications of materials under study, with an application focus.

Abstract Patients with a neuropathic bladder have chronic medical problems with urinary incontinence, urinary infections, and potential renal failure. Conventional surgical management of the neuropathic bladder uses detubularized bowel as a patch (enterocystoplasty) to enlarge the bladder. However, this structural modification provides no functional improvement, and carries other complications. Alternative methods to enterocystoplasty have been explored through tissue engineering, by regrowing cells on biodegradable polymers or decellularized biological matrices. In all cases, some elements of regenerated bladder tissue structure have been obtained, but full bladder function has not been restored. Recent advances in nanotechnology provide a novel and alternative strategy for the use of tissue engineering scaffolds. Peptideamphiphile (PA) biomaterials have been developed, which self-assemble on the nanoscale to form fibrous gels, capable of retaining and releasing critical growth factors for regeneration. By non-covalently binding growth factors to the gel matrix, they are protected from degradation and can be released over time. The hypothesis of this proposal is that controlled delivery of bFGF and VEGF from a self-assembling nanofiber PA -scaffold composite will enhance the regeneration of bladder tissue. In this proposal, two well established complementary methods of binding growth factors to PAs will be characterized for their ability to effectively bind and release bFGF and VEGF (Aim 1). The in vitro effect of these PAs will then be assessed by seeding bladder cells within PA-scaffold composites and determining the effects on cell proliferation and differentiation (Aim 2). Lastly, the effects of the PAs on tissue regeneration will be studied in vivo by seeding bladder cells on PA-scaffold composites in a nude rat model. Differences in smooth muscle formation and angiogenesis in the regenerated tissue will be determined (Aim 3). This proposed research will provide the first evaluation of a novel tissue engineering nanotechnology for bladder regeneration

Award # 1R41DK074254-01 Agency NIH Lead Institution EMV Technologies, LLCTypeExtramural

Explanatory Notes The new kidney dialysis membranes under study in this project will be characterized for parameters including those that affect the chemical or physical properties of the material. These metrology efforts support the ability to understand the effects of modifications to nanomaterials on their properties.

Abstract End stage renal disease (ESRD) has consistently been a major medical problem worldwide. By the year 2010, the number of ESRD patients in the United States is projected to be 661,330 with an increase of 78% compared to 372,407 in the year 2000. The total annual cost of Medicare for ESRD patients in the United States is projected to be $28.3 billion by the year 2010, an increase of 99% compared to $14.2 billion in the year 2000. Current hemodialysis membranes used in kidney dialysis have difficulty in removing uremic toxins from the blood. Nano-porous alumina tubes have gained significant interest in drug delivery and gas separation technologies and, as of yet, their use as membranes for hemodialysis has not been researched. The objective of this project is to create nano-porous alumina tubes with average pore sizes of 5 nm using a process of anodizing aluminum tubes under controlled conditions. There are several advantages of alumina membranes over the membranes currently used including: high porosity and uniform pore size, high hydraulic conductivity (water permeability), uniform pore distribution, excellent pore structure, high resistance to chemical and thermal degradation, and superior mechanical properties. The pore size and porosity will be measured by Scanning Electron Microscopy (SEM), while membrane permeability will be tested by a custom designed setup. Finally, a prototype mini-module hemodialyzer will be constructed and the clearance of a wide spectrum of molecular weight solutes (from low to high molecular weight solutes) will be evaluated. This research will address the needs of the National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK), specifically its Division of Kidney, Urologic, and Hematologic Diseases. The NIDDK is seeking "the development of new dialysis membranes to diminish the duration of dialysis treatments." When commercialized, the new membranes will deliver a higher average dose to dialysis patients and increase their quality of life

Award # 5P20MD001085-02 Agency NIH Lead Institution Albany State UniversityTypeExtramural

Explanatory Notes This project supports efforts to understand and characterize nanoparticle surfaces and modifications to such surfaces.by using nanotechnology to develop the basic science of nanoparticle thin films.

Abstract Albany State University (ASU) is proposing to improve its biotechnology infrastructure to enhance its research capabilities. In the proposed RIMI program, we intend to include three capacity building components that deal with the research infrastructure, faculty development and collaborative research. Although ASU has built a good research infrastructure it lacks well structured research facilities in biomedical sciences. We seek to develop research facilities to include a core biotechnology lab, tissue culture facility, common reagent room, and a bio-imaging facility. We have incorporated a strong faculty development component after well-planned and thoughtful discussions with our faculty. This includes release time support, on-campus/off-campus research support, a biomedical seminar series, technology training workshops, and scientific meeting support. The collaborative component was added to broaden our existing collaborative efforts with Georgia Institute of Technology, Tennessee University and Purdue University. A central research support service is proposed in the RIMI program which will be an administrative unit coordinating the efforts of the RIMI program and managing program's central resources and activities. The RIMI program will play a major role in providing resources and infrastructure at ASU to improve faculty productivity in terms of proposal writing, scientific presentations and publications in referenced journals. With the collaborative research projects, the research environment at ASU will be enhanced which will result in increased minority student participation in biotechnology research and an increased minority pool of biomedical researchers

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program supports investigations which seek to understand the biological or environmental response of a nanomaterial to changes in a defined set of physical parameters.

Abstract Mathematical modeling that facilitates estimation of the biological or environmental response of a nanomaterial to changes in a defined set of physical parameters is an important element for assessing environmental, health, and safety aspects of nanomaterials. NIST has several programs in data and informatics research in support of nanomaterial modeling. One example concerns the magnetic properties of metallic nanoparticles relevant to the use of metal nanoparticles for treatment of cancer and hyperthermia and as medical imaging contrast agents and drug delivery systems. Such modeling may prove useful for facilitating fabrication of nanostructures that are better and safer through design

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes Nanomaterial characterizations of direct relevance to the standardization of toxicology protocols are an intergral component of this program.

Abstract The three gold RMs are a direct result of a January 2006 request by the Office of the Director of the National Cancer Institute (NCI) for a series of nanosize gold-based particle standards. They are tailored to meet the needs of the bio-nanotech community and specific toxicological protocols for nanomaterials being developed at NCI. The materials are also critically relevant to the standardization process undertaken by NCI-NIST-FDA partnership in collaboration with the National Toxicology Program and National Environmental and Health Institute.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program seeks to provide measurement tools for tissue engineering research. High-throughput and combinatorial methods for characterizing biomaterials and screening cell-material interactions are large areas of investigation.

Abstract The goal of regenerative medicine is to use materials and signaling molecules to guide cells in repairing damaged tissue. The large parameter space that must be considered in developing these therapies makes combinatorial approaches attractive. The problem is particularly complex because both composition and processing variables have to be considered. We are developing gradient libraries – single samples containing a continuous variation in material parameter along one or two orthogonal directions – as a measurement tool for tissue engineering research. This project will develop high-throughput and combinatorial methods for characterizing biomaterials and screening cell-material interactions. There will also be combinatorial libraries to serve as reference materials (test patterns) and genetically engineered cells capable of fluorescently indicating their state in real time (indicator cells).

Award # Agency DOE Lead Institution University of TexasTypeExtramural

Explanatory Notes Quantification of mass transport through single-nanopore models having well-defined structures is important to understanding process of technological and envronmental significance, and supports the development of methods for the determination of particle size.

Abstract The goal of this project is to quantify mass transport through single-nanopore models having well-defined structures. Our specific objectives in this proposal are to measure the hydrodynamic hindrance factors in a carbon nanotube channel and to study the effects of probe-pore interactions on transport rate.
Quantifying mass transport through nanoporous media is important, because it is the foundation for many technologically significant processes including separations, analysis, and catalysis. Mass transport through nanopores involves restricted motion (hindrance) of molecular and larger probes when their sizes are comparable to the pore size; thus hindered transport is of particular interest when the pore size is on the nano scale. For example, understanding hindered transport is critical for developing an understanding of the selectivity of catalytic reactions in zeolites and size-exclusion-based separation techniques. In addition, chemical interactions between probes and pores are crucial for zeolite-based catalytic reaction and nanopore-based chemical sensing.
There have been many studies of hindered transport that have involved array-pore membrane models, but only few using single-pore models. This is largely a consequence of the difficulty associated with quantifying transport through a single, well-defined nanopore using conventional analytical methods. However, during the previous grant period we developed a sensitive and robust method for studying stochastic transport of nanometer-scale particles passing through the pores of individual multiwall carbon nanotubes. Our approach studying transport avoids most of the drawbacks of many-pore models, which include non-uniform pore size and shape distribution and scaling by equilibrium partition, and significantly improves upon existing experimental models for studying single-pore transport.

Award # 1R01OH008807-01 Agency NIOSH Lead Institution New York University School of MedicineTypeExtramural

Explanatory Notes

Abstract Carbon nanotubes (CNT) are dominant among the array of nanomaterials because of their unique chemical and physical properties. Promising applications in many areas are expected to lead to industrial scale production in the near future. CNTs could become airborne during manufacturing and handling and result in inhalation and dermal exposure of workers to particles of unknown toxicity. However, knowledge is limited regarding potential exposure concentrations for workers exposed to this new type of material. Also, no adequate method exists for quantitative and qualitative monitoring of airborne CNTs because of their complexity. The proposed research will develop a comprehensive yet practical method for sampling, quantification, and characterization of CNT particles in air. The method will permit classification of sampled particles into three categories: tubes, ropes (bundles of single-walled CNTs bounded by Van der Waals attraction force), and nontubular particles (soot, metal catalysts, and dust, etc.). The method will also permit calculation of the number concentrations and size distributions for each type, and the shape characters (diameter, length, aspect ratio and curvature) of CNTs. The method will use available instrumentation to build an air monitoring system that is capable of sampling and sizing airborne CNT particles in a wide size range by using a 10-stage Micro-orifice uniform Deposit Impactor (MOUDI) and an Integrated Diffusion Battery previously developed in this laboratory. The samples of each size fraction will be collected onto Silicon-chip substrates and analyzed using Atomic Force Microscopy (AFM). Newly developed software, SIMAGIS® Nanotube Solutions, will be used for AFM image analysis and data processing, which can automatically count nanotubes, nanoropes and particles; and measure the shape characters. Other commercially available nanoparticle sampling instruments, such as an electrostatic aerosol sampler and a Nano-MOUDI will also be tested in this work. Successful completion of this project will produce a validated method for sampling airborne CNTs in the workplace and a practical method (using AFM image analysis technology) for classifying sampled CNT particles by type and for quantifying and characterizing each type separately. These methods are needed for determining health risks that may result from worker exposure to the various types: CNTs, nanoropes, and nontubular nanoparticles. The results will also provide a foundation for field and personal sampling devices for CNTs.

Award # Agency NIST Lead Institution NISTTypeExtramural

Explanatory Notes This program supports the development of techniques to monitor and characterize airborne nanoparticles.

Abstract

Award # 0526977 Agency NSF Lead Institution University of Arkansas Little RockTypeExtramural

Explanatory Notes This effort is directed toward the development of an instrument to study the properties of minute-sized airborne particles.

Abstract The University of Arkansas at Little Rock (UALR), in collaboration with Arkansas State University, Arkansas Children's Hospital, and the University of Arkansas for Medical Sciences, is proposing to develop an instrument to study the properties of minute-sized airborne particles and their roles in environmental health, science, medicine, and economic development. These particles, too small to be visible even with the aid of a standard laboratory microscope, may cause serious damage to both the lungs and the heart (Science Magazine, 25 March 2005), and these adverse health effects range across many industries - from particles present in soot and automotive exhaust to lunar or Martian dust that astronauts will be exposed to in human missions to the Moon and Mars. In the emerging development of nanotechnology, industries thus face many challenges in controlling the hazards presented by nanoparticles, and perhaps even more importantly are the myriad beneficial applications to engineering and medical technologies that nanotechnology can provide. These nanoparticles may be assembled together by non-contact forces in a precise structure and order to form new engineering materials and medical drugs not feasible for assembly with any current mechanical tools in existence. This, for instance, opens the pathway to new drug development and delivery techniques for cystic fibrosis patients. Experts predict that economic development to address the two sides of currently emerging nanotechnology will grow into a trillion dollar industry in the United States alone over the next two decades. UALR will develop a new instrument to measure the size and static charge distribution of the particles and to control their motion in electric, magnetic, and gravitational fields. Under the proposed two-year instrument development application project, it will be used to study: 1) Material coatings development for medical devices, and new drug development for respiratory delivery, 2) studies on the adverse environmental effects of nanoparticles from various industries, 3) detection of chemicals and biotoxins in the air for US Space and Missile Defense Command, and the development of new respiratory protection against nanoparticle inhalation, and 4) the improvement of advanced engineering processes in reducing the emission of nanoparticle exhaust pollutants in the automotive industry. The proposed research is designed to support integrative education of students from grade 9 up to the completion of a PhD program, as the university faculty members will work with school teachers preparing the students for education and research at the university level.Technical Abstract: Development of a laser based instrument to measure simultaneously both particle size and electrostatic charge distributions in real time and on a single particle basis for particles in the size range 10 to 1000 nm in diameter is proposed by a team of researchers of the University of Arkansas at Little Rock (UALR). The instrument will employ a Laser Doppler Velocimeter (LDV) that will analyze the response of particle motion under the excitation of ac electric and acoustic fields synchronously applied at a frequency ranging from 10 to 100 KHz. The diameter (da) and the charge (q) of each particles passing through the LDV sensing volume, are determined by measuring the phase lag and the amplitude ratio of the oscillatory particle motion with respect to the applied sinusoidal fields. A high frequency photon correlator and a cooled photomultiplier tube will be used to process signals from the radiation scattered from particles to measure the size and charge distributions at a rate of 1000 particles per second. Application of this new Electro-acoustic Single Particle Aerodynamic Relaxation Time (ESPART) Analyzer for nanoparticles will be utilized in several research projects being performed by the team of scientists, engineers, and physicians at UALR, and other campuses in the US and abroad including Arkansas Children Hospital and the University of Arkansas for Medical Sciences. These projects include: 1) Material engineering studies of aerodynamic and electrokinetic transport of nanoparticles, their deposition and cluster formation, electrodynamic guided assembly of airborne particles of different sizes and compositions developing new drugs for respiratory delivery, and electrospray of nanoparticles for coating cardiovascular stents 2) studies on the regional lung deposition of inhaled nanoparticles as a function of size and charge using physical models, 3) detection of chemicals and biotoxins in air for US Space and Missile Defense Command, and the development of new respiratory protection against nanoparticle inhalation, and 4) plasma processes in reducing the emission of nanoparticle exhaust pollutants in the automotive industry. The proposed research and education project is designed to support integrative education of students anywhere from grade 9 to the completion of a PhD program in material engineering, as the university faculty members will work with school teachers and students in preparing the students for education and research at the university level

Award # 1R01DK073991-01A1 Agency NIH Lead Institution Georgia Institute of TechnologyTypeExtramural

Explanatory Notes This project focuses on the development of tissue engineered substitutes with an emphasis on understanding the effects of cryopreservation on cells and biomaterials. This is critical groundwork for the development of cell- or bio-based reference materials.

Abstract Tissue engineered substitutes have the potential to provide effective, functional replacements of failing tissues and organs and as such help resolve the current transplantation crisis. To realize its potential, tissue engineering must generate substitutes that can be preserved in the long-term. Preservation is essential for off-the-shelf availability, storage and distribution of constructs fabricated at a large-scale at centralized locations, as well as for sterlity testing and quality control. The long-range goal associated with this research program is to develop a fundamental understanding of how the various cryopreservation parameters affect cellular and tissue construct function, and on the basis of this knowledge to develop widely applicable preservation protocols. The objective of this application is to evaluate the effect of ice-forming and ice-free cryopreservation on cell viability and construct function for a model pancreatic tissue substitute consisting of insulin-secreting cells and a hydrogel biomaterial. The central hypothesis is that entire tissue engineered substitutes can be successfully vitrified, while maintaining key structural and functional features of the cellular and biomaterial components; vitrification produces superior outcomes relative to conventional, ice-forming cryopreservation in terms of both construct integrity and functionality. Guided by strong preliminary data, the hypothesis will be addressed by the following 3 specific aims: 1) determine cell osmotic tolerance limits and cryoprotectant cytotoxicity and define the domain of conditions under which to cryopreserve cells used in tissue substitutes; 2) characterize in vitro the effect of cryopreservation on construct structure and function; 3) evaluate the in vivo functionality of cryopreserved substitutes. The effect of cryopreservation on cells will be assessed on the basis of cell viability, apoptosis, stress protein expression, and metabolic and secretory functions; and on biomaterials on the basis of their structural integrity and functionality. The approach is innovative, as it focuses on studying the effects of cryopreservation on both the cellular and biomaterial components of a three-dimensional (3D) tissue substitute. The proposed research is significant, as it will generate new fundamental information on the science of cryopreservation, provide a quantitative framework for the rational design of cryopreservation protocols, and identify key design features of tissue substitutes that enable their cryopreservation.

Award # 5R43ES013367-02 Agency NIH Lead Institution LNKChemSolutionsTypeSBIR

Explanatory Notes This research is dedicated to the manufacturing of nanoparticulates for toxicology research. Such research supports the development of standard materials and standardized toxicological testing protocols.

Abstract This Phase I SBIR program is aimed at initiating a business venture dedicated to the manufacturing of nanoparticulates for toxicology research. The rate of nanotechnology R&D growth in the commercial, academic, and private sectors, appears to be outpaced only by the increasing level of concern among policy makers, Federal agencies, academic researchers and the general public regarding the toxicity of new nanomaterials. We are convinced that our initiative is timely, and that it responds a true societal need. The proposed business venture is based on a unique set of manufacturing methods to design a wide variety of well-defined nanostructured powders, and a partnership with the Midwest Research Institute is established to integrate toxicology tests with the nanomanufacturing component, and into a business plan involving professionals with diverse backgrounds. The proposed effort also involves participation of Kamterter II, LLC, another Lincoln, NE based small business with an interest in nanotechnology applications for the agrochemical industry. LNKChemsolutions will contribute the know-how for nanomaterials synthesis. The MRI has code-compliant barrier facilities to handle the more toxic samples, and a proven track record of research with animal models. Kamterter II, LLC, provides a platform to target the agrochemical secdtor, who is becoming increasingly more concerned with the formulation, use, and environmental impact of pesticides in their nanoparticulate forms. Preliminary toxicology tests on three model systems with real commercial potential, and the design of such nanostructured materials, will serve as a platform to launch a spinoff business venture dedicated to serving the health sciences community and the private sector concerned with the environmental and occupational health impact of new nanotechnology products. The three case studies will include nanoparticle synthesis and preliminary toxicological testing of one metal oxide, one pesticide, and one multiactivity phytochemical. On completion of Phase I research, we expect to appeal to a broad clientele for custom synthesis of well-defined and characterized nanomaterials for toxicological evaluation, or for contract research and testing

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program targets the development of specific reference materials for nanomaterials.

Abstract A carbon nanotube Reference Material is under development at NIST. Reference Material 8475 Carbon Nanotube Bearing Material is a product of commerce that constitutes an as-received sample from the manufacturer. The material contains single-walled, closed-ended, carbon nanotubes. The material was produced by an arc method in a single process run. The material was packaged into 0.35 g to 0.5 g units and was recently used as part of an interlaboratory pilot study to characterize nanoparticles for properties such as surface area. In addition, the material was provided to NIOSH as part of an effort that includes collaboration between NIST, NASA, NIOSH, NRC, University of Maryland, and Lehigh University for nanomaterial characterization research in support of assessing environmental, health, and safety aspects of nanomaterials. Reference materials, such as the carbon nanotube material described above, provide the foundation for assessment of analytical chemistry methods, accurate quantification of occupational and environmental exposures, and are useful for health effects research via in vitro and in vivo toxicology. NIST supplies industry, academia, government, and other users with over 1300 reference materials of the highest quality and metrological value, and a number of these are useful for assessing environmental, health, and safety of nanomaterials.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program targets the development of specific reference materials for nanomaterials.

Abstract Standard Reference Materials (SRM) and Reference Materials (RM) have been developed for identifying and quantifying asbestos types, which are naturally occurring nanoparticles with significant and well known nano-EHS impact. These reference materials include: SRM 1866a, SRM 1867a, SRM 1868, and SRM 1876b and RM 8411. The last two of these are intended for evaluating the techniques used to identify and count fibers by transmission electron microscopy, critical for quantitative understanding of nano-EHS impact. Several particle size standards in the nanometer to micrometer size range are available. These are useful for evaluating or calibrating specific types of particle size measuring instruments such as electrical zone flow-through counters, optical and scanning electron microscopes, light scattering instruments, sedimentation systems, and wire cloth sieving devices. Many of these materials are polystyrene particles that consist of commercially manufactured monodisperse latex particles in water suspensions. The smallest of these in terms of particle size is a monodispersed polystyrene material at 60 nm. This material will be very useful for assessing clean room quality in the electronics industry and will be important for accurately sizing of particles in this size range. They are particularly critical for calibrating surface scanning inspection systems. The reference particles may also be used for supplying monosized particles for testing aerosol instruments and are useful for examining aerosol kinetics and evaluating particle detector response for airborne chemical and biological agents.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program targets the development of specific reference materials for nanomaterials.

Abstract There is a comprehensive metrology program focused on the three critical interrelated issues facing carbon nanotube applications: quality, characterization, and alignment. This program will enable NIST-Quality measurements on this important material that are impossible without high quality, well-characterized samples. Success will accelerate development across a broad range of anticipated applications through standard measurement methods facilitating commercial interaction and a knowledge base of materials properties. Moreover, this program will build critical skills in several areas, setting the foundation of future NIST programs. The separations work extends a well-honed set of experimental capabilities developed for polymeric materials into new materials classes. The interwoven relationships between spectroscopy and the theory will be applied to increasingly complex systems. Additionally, there will be an increase in NIST expertise in phage display technology, a powerful new bio-inspired approach to characterizing the interface between inorganic and organic materials.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This program targets the development of specific reference materials for nanometrology via scanning probe microscopy (SPM).

Abstract Accurate and validated protocols and reference materials will be developed to define the uses and limitations of major analytical methods.

Award # Agency DOE Lead Institution Ames LaboratoryTypeExtramural

Explanatory Notes This program seeks the development of new methodologies to advance mapping of surfaces in the nanoscale environment.

Abstract New methodologies in analytical chemistry and separations science are being developed to address advances in heterogeneous and homogeneous catalysis, nanotechnology, biomimetic systems, environmentally-benign chemistry and toxic waste clean-up. Microenvironments on surfaces are being characterized to provide fundamental understanding about catalytic and separation schemes, including mapping surface heterogeneities and studying interactions at microstructures. To address key characterization issues in the next decade, the Ames team will focus on the development of new concepts for chemical analysis and the evaluation of structure-function relationships at nanometer length scales and in microenvironments. These projects range from characterizing reactions at solid/gas and solid/liquid interfaces to probing details of heterogeneous chemical reactions and of chromatographic processes. By drawing on the unique expertise of the individual groups through collaboration on these and other projects, the team is well positioned to play a major role in the National Nanotechnology Initiative. The results of these studies are expected to impact other areas of interest to DOE as well, such as separations, catalysis, energy conversion, and environmental monitoring.

Award # Agency DOE Lead Institution Argonne National LaboratoryTypeExtramural

Explanatory Notes This program supports the development of advanced electron microcopies for atomic scale characterization of spatio-chemical composition.

Abstract The goal of this program is to develop atomic-scale three-dimensional chemical imaging of interfaces in nanostructures to gain fundamental knowledge of the chemical origins of their novel nanoscale properties. This is accomplished via the ground-breaking use of a combination of electron tomography, energy-filtered transmission electron microscopy (EFTEM) and aberration correction. This approach, based on energy filtered imaging, allows faster data acquisition of larger areas as compared to STEM techniques. The combination of EFTEM and aberration correction will enable the field of 3-D reconstruction to evolve beyond chemical mapping at the 1nm scale to elemental mapping with atomic resolution and to mapping of chemical bonding. Aberration correction has the potential to improve the resolution in energy-filtered imaging, which is limited up to date to ~1nm for many applications, to the atomic level. This combination of methods will improve resolution, sensitivity and validity of data interpretation beyond anything that is currently available.

Award # Agency DOE Lead Institution Argonne National LaboratoryTypeExtramural

Explanatory Notes This effort explores the interactions of energetic particles with nanoscale surfaces. Analysis of such information can provide mass-based analysis of nanomaterial and detailed information on trace-level impurities.

Abstract The interaction of directed energy sources such as energetic ions, electrons, and photons with surfaces provides the basis for modifying, patterning and analyzing surfaces and nanoscale materials. This program investigates the fundamentals of these complex interactions over a wide range of conditions using several unique, world-class methods developed in our laboratory. These uniquely sensitive tools for trace analysis are also providing, for the first time, mass based analysis of materials with nanometer scale lengths.

Award # Agency DOE Lead Institution Brookhaven National LaboratoryTypeExtramural

Explanatory Notes This work will document property-sensitive nanoscale structure and defects of materials through a combination of experiment, modeling, and simulations.

Abstract The goal of this program is to study property sensitive nanoscale structure and defects in technologically-important materials such as superconductors, magnets, and other functional materials. Advanced quantitative electron microscopy techniques, such as coherent diffraction, atomic imaging, column-by-column spectroscopy, and phase retrieval methods including electron holography are developed and employed to study material behaviors. Computer simulations and theoretical modeling are carried out to aid the interpretation of experimental data. Fabrication of thin films with tailored microstructure and nano-assemblies to understand materials’ electronic and magnetic behaviors is also incorporated.

Award # Agency DOE Lead Institution Lawrence Berkeley National LaboratoryTypeExtramural

Explanatory Notes This program aims to develop novel microscopies for the analysis of nanostructured materials, including methods for assessing purity and heterogeneity of nanomaterials.

Abstract This program aims to stretch the limits in the state-of-the-art of optical characterization, through the development of novel forms of optical microscopies, such as single pulse CARS microscopy with phase control, aperture-less NSOM (ANSOM), and femto-second pump-optical injection probing spectroscopies of single nanostructure species under stimulated emission conditions. A parallel molecular beam epitaxy growth effort for processing semiconductor materials with in situ scanning tunneling microscopy is used to study the growth mechanisms of III-V nitrides and doped nitrides. Development of novel microscopies provide new spectral and spatial windows into the analysis of materials for DOE missions in energy utilization.

Award # Agency DOE Lead Institution University of California-Los AngelesTypeExtramural

Explanatory Notes This program supports the development of methods to determine the 3D structures of single particles down to the atomic level.

Abstract Visualizing the arrangement of atoms has played a crucial role in understanding the microscopic world. There are already a few ways of imaging atomic structures, but each has its limitations. Scanning probe microscopes are limited to imaging atomic structures at the surface. Transmission electron microscopes can resolve atoms but only for samples thinner than ~ 30 nm. Crystallography can reveal the globally averaged 3D atomic structures based on the diffraction phenomenon, but it requires crystals. These limitations can in principle be overcome by coherent diffraction imaging that is based upon the principle of using coherent x-ray scattering in combination with a method of direct phase recovery called over-sampling. Coherent diffraction imaging has been successfully applied to 2D and 3D imaging of nanoscale materials and biological samples. A highest spatial resolution of 7 nm has been achieved, while the ultimate resolution is only limited by the X-ray wavelengths. By using the 3rd generation synchrotron radiation sources, better-designed instruments and more robust image reconstruction algorithms, we expect to improve the spatial resolution to the 1 nm level within the next few years. Meanwhile, we will also pursue its applications in materials and nano science. We will focus on 3D imaging of porous silica particles, GaN semiconductors, quantum dots and mineralized bone. The ability to image the internal pore structures in three dimensions, 3D morphology of GaN and its alloys in nanocrystal form, and 3D internal structures of quantum dots, coupled with computational methods such as molecular dynamics and ab initio calculations, will profoundly expand our understanding of the critical structural and morphological features required to make superior catalysts, adsorbents, electrodes or semiconductors. Understanding the mineral component of bone such as the size, shape and arrangement of the calcium apatite crystals in a collagen matrix will be of fundamental importance in biology and medicine. In the long run, with the prospects of brighter X-ray sources and pixel array detectors of higher quantum efficiency and a higher dynamic range, coherent diffraction imaging could potentially be used to determine the 3D structures of single particles down to the atomic level.

Award # Agency DOE Lead Institution Harvard UniversityTypeExtramural

Explanatory Notes These structural studies support the development of methods to assess dimensional and chemical properties of nanomaterials.

Abstract The current research program extends electron diffraction measurements to small metal clusters which have shown interesting properties as nanocatalysts. These diffraction techniques have achieved the capability to directly observe the evolution of cluster structure with size and temperature for sizes in the range of 10 – 100 atoms. In particular, one of the research goals is the detection of changes in structure induced by adsorbed species which can provide important data to help form a theoretical model of the nanocatalytic process. Recent measurements of low energy structural isomers of silver clusters have identified that short range order having fivefold symmetry dominates many of these structures in the size range <50 atoms; a result that has been shown to be consistent with structures derived from density functional calculations. These results provide an appreciation for the complexity and richness of cluster structures and introduce new methods to study the science of disordered materials, possibly leading to novel applications of nanostructure materials to catalysis. The research program will investigate the size and temperature dependence of gold and silver clusters to identify trends of structural order; specifically, the appearance of local symmetry having fivefold order as well as globally ordered clusters. The analysis of cluster structures with short range order will be related to the more general understanding previously developed for bulk metals. The structures of both gold cluster cations and anions will be measured to determine the predicted 2D to 3D transition and to characterize structures in the size range of 10-30 atoms which have been identified as catalytically active. Questions related to the rearrangement of cluster atoms upon the adsorption of oxygen molecules will be investigated by measuring the diffraction of gold clusters with adsorbed oxygen. The temperature and cluster size dependence characterizing the equilibrium adsorption of oxygen molecules on gold cluster anions has been studied recently in our laboratory. The structures of clusters which have previously demonstrated an enhancement of catalytic activity in the presence of water vapor will also be studied. The diffraction of bimetallic clusters of catalytic interest will be performed. Although these materials have been well studied for bulk materials, they have not been considered previously in cluster studies. The nanocatalyst candidates include bimetallic clusters of iron-gold, nickel-gold and silver-gold. Diffraction measurements of such cluster materials will be obtained for non-stoichiometric compositions formed by a sputter aggregation source. The diffraction dependence on bimetallic composition will help to understand how the interplay of structure and oxygen adsorption depends on composition. Species identified as interesting catalytic substrates, for example by level of adsorption and structural changes, will be further investigated and correlated with theoretical models. These diffraction measurements will rely on the availability of density functional calculations to provide isomer structures with which to compare data. Several theoretical groups are collaborating on current research projects and will continue to be a source of support for these planned measurements.

Award # Agency DOE Lead Institution University of Illinois-Urbana-ChampaignTypeExtramural

Explanatory Notes This programs seeks to develop techniques to determine the structures of individual nanostructures and complex crystals, ultimately with atomic resolution in three dimensions.

Abstract Determining the atomic structure of complex molecules such as proteins is typically performed using single crystal X-ray or neutron diffraction techniques. For polycrystalline materials, powder diffraction methods can be utilized, but information obtained is sampled over many grains and provides only the averaged crystal structure. Both X-ray and neutron diffraction require perfectly homogenous materials. Many non-periodic structures found in nature and used in technology are either too complex or too small for these methods; their structure consequently has not been accessible to crystallographic techniques. Since the properties of materials ultimately depend on their local atomic structure, understanding structure-property relationships in nanomaterials thus depends critically on further progress in nanostructure characterization. In the initial phase of this DOE grant, we have developed a new nanometer scale structural characterization technique. This technique uses the formation of a nanometer-sized, highly coherent and parallel, electron probe to record quantitative diffraction patterns from individual nanostructures. Here, we propose to further develop this electron crystallographic technique for scanning nanoarea electron diffraction and diffraction based 3-D, atomic resolution, tomography, and apply the techniques to the structural determination of individual nanostructures and complex crystals. The proposed research takes advantages of the spatial resolution offered by a nanometer-sized coherent electron probe and the quantitative diffraction information obtained from the high angular resolution achieved with a parallel illumination. We will approach the challenging problem of 3-D atomic resolution tomography by using a combination of high-resolution electron diffraction and ab initio phase retrieval for image reconstruction. The advantage of this approach is that the resolution is only limited by the signal and noise ratio of high frequency information in the diffraction patterns rather than the electron lens aberrations. The advanced electron diffraction techniques will be applied to determine local structures of real crystals, selected nanocrystals and multi-wall tubes. Special focus will be on multi-phase functional oxides, for which the lack of local structural probes has limited our understanding of their properties. The electron diffraction techniques developed here are general, applicable to a broad range of nanomaterials and can be implemented in many existing electron microscopy facilities across the country. Broad impact is expected from the contribution of this valuable new characterization tool to the nation’s science and technology programs. Broad impact is also expected from advances in understanding of nanoscale materials garnered from the applications of this technique.

Award # Agency DOE Lead Institution Stony Brook UniversityTypeExtramural

Explanatory Notes This work will improve and standardize data and image handling for techniques three dimensional imaging of nanostructures.

Abstract Our efforts are focused on exploring some of the fundamental issues in diffraction microscopy. With Veit Elser’s group at Cornell University, we are looking at how to define resolution, and obtain the best reconstruction. We are establishing file formats that we propose as standards to the community, and a willingness to share our code. We are looking into the issues of how best to merge 2D data into 3D arrays. We are considering the issues of optimum wavelength for a dedicated beamline for diffraction imaging at the Advanced Light Source.

Award # 1R43DK074237-01 Agency NIH Lead Institution Miniature Tool And DieTypeExtramural

Explanatory Notes This project measures flow in nano-volumes, furthering the evaluation of the physical properties of materials at the nanoscale, to support metrology needs and the development of methods to characterize a material's composition, purity, or heterogeneity.

Abstract Currently, there is a serious disconnect between macro and micro injection molding industry. Manufacturers' polymer material data sheets that predict the physical and mechanical properties including flow do not predict properties below 0.020" thickness.. Since properties have proven to be non-linear in thin-walled applications, data below 0.020" may not be able to be accurately predicted. Although there are numerous ASTM standards for material property characterization of macro injection molded parts, none exist for micro- molding. The goal of this proposal is to predict the flow of polymer materials into micro cavities with nano volume features. This information will accelerate the design of thin-walled (<0.010") micro injection moldable parts for numerous commercial applications. In order to achieve this goal we will start by creating standards to evaluate the physical properties of materials with wall thicknesses of <0.010". These same standards would be useful in characterizing new enhancements or processes in material science. In Phase I, we will begin to investigate and characterize one such process. We will compare "neat" to "treated" polymer performance in the production of thin-walled micro injection molded samples

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes This development of methods to determine the distribution of chemical species in three dimensions directly supports this research need.

Abstract This program will develop a quantitative understanding of the distribution of chemical species in three dimensions, including the internal structure, interfaces, and surfaces of micro- and nanoscale systems. These approaches will be broadly applicable to nanoscale technologies from microelectronics to pharmaceuticals and subcellular biomedical applications. The ultimate analytical challenge is the cell and combinations of cells in biosysems. These are the most chemically complex nanodevices. Small chemical changes in these 3D-complex systems (within and between cells) make these biodevices work. Understanding and imaging these chemical changes are critical to many biomedical applications.

Award # Agency NIST Lead Institution NISTTypeIntramural

Explanatory Notes Measurement tools are being developed to characterize size, shape, molecular orientation, defect structure, and the elastic or anelastic properties needed to determine the stability of model nanostructures.

Abstract Applications of nanotechnology will fail in the absence of robust nanostructures. To enable versatile and successful nanomanufacturing methods, metrology must be developed to correlate the nanoscale materials properties with processing and performance. This project uses nanoimprint lithography (NIL) to manufacture structures with systematic variations in dimensions, structure, and materials properties. Measurement tools are developed to characterize size, shape, molecular orientation, defect structure, and the elastic or anelastic properties needed to determine the stability of model nanostructures.

Award # 0449268 Agency NSF Lead Institution SUNY at Stony BrookTypeExtramural

Explanatory Notes This work is targeted to obtaining small samples from the surface of nanostructured materials or nanostructured particles in order to characterize them and obtain spatial distribution of the nanomaterial properties.

Abstract This research plan involves cross-disciplinary applications of instrumented indentation to the investigation of elastic-plastic deformation in a number of inorganic and biological materials systems of increasing visibility and fundamental importance to the engineering and physiological communities, and spanning nano- to macro-scopic size scales. The power of indentation as a scientific tool lies primarily in its experimental simplicity, due to the minimal specimen preparation involved. However, interpretation of results is non-trivial and key to successful analysis are sufficient analytical modeling and supplementary observation. In this plan, indentation will be used to develop constitutive behavior of three 'testbed' systems: i) rapidly quenched small-volume structures on substrates, ii) carbon nanotube arrays on substrates, and iii) lung parenchyma. Academic, industrial and/or clinical collaborators have been identified to add perspective and disseminate information. Efforts are planned to develop new interdisciplinary research courses for university, and practical and pre-collegiate training programs for K-12 students.

Award # 0619398 Agency NSF Lead Institution University of California-MercedTypeExtramural

Explanatory Notes This diffractometer will be used for qualitative and quantitative phase identification, amorphous/crystalline characterization, polymorph discrimination, and impurity analysis in envronmental and materials research, including further development of methods to study nanoparticles and their properties.

Abstract The acquisition of a powder X-ray diffractometer (XRD) system serves a variety of research and teaching needs at UC Merced, the new 10th campus in the UC system, which officially opened its doors for undergraduate and graduate students in Fall 2005. Powder XRD is an essential tool for ongoing environmental and materials research, and a critical instrument for the expansion of our interdisciplinary materials science and engineering and chemical sciences programs. We are purchasing a basic, high quality instrument with automated capabilities that can be easily reconfigured for different applications to serve a range of users and research demands. Features of the instrument include a solid-state X-ray generator with Cu X-ray tube, theta-theta goniometer (for a stationary sample, moving source and detector), fast solid-state detector with low background, and a temperature-controlled, inert gas sample chamber attachment (-193 C to +450 C) for air-sensitive samples and investigation of temperature-dependent phase changes. The XRD instrumentation will be openly available to users as part of our existing Imaging and Microscopy Facility, the first centralized campus recharge facility, and supported by a full-time laboratory manager who will be responsible for instrument oversight and user training. It will support faculty, graduate, and undergraduate research, and it will be used in courses such as environmental chemistry, instrumental/analytical methods, soil chemistry, materials science, and materials engineering. Current and planned research that will be supported at UC Merced includes: the chemistry and mobility of contaminants in the environment; environmental chemistry and mineralogy of geoparticles in soils, sedimentary environments, and engineered systems; atmospheric and environmental agglomerates and nanoparticles; properties of natural biomaterials; synthesis, characterization, structure, and properties of semiconductor nanoparticles. Main applications include qualitative and quantitative phase identification, amorphous/crystalline characterization, polymorph discrimination, impurity analysis, and temperature- and atmosphere-dependent phase transformation studies. The instrumentation will have a broad impact in contributing to UC Merced's mission to serve the rapidly growing population of the southern Central Valley of California, which is dominantly Hispanic, has a high immigrant and first-generation population, and is historically underrepresented in higher education.

Award # 0310717 Agency NSF Lead Institution Colorado State UniversityTypeExtramural

Explanatory Notes This project focuses on the goal of making EUV light, now mostly limited to a handful of large national facilities, available routinely in a broad variety of laboratory settings, for applications such as high-resolution imaging, spectroscopy, elemental- and bio-microscopy, and nano-fabrication.

Abstract Welcome to the Center for Extreme Ultraviolet Science and Technology – an Engineering Research Center (ERC) exploring the development of compact coherent extreme ultraviolet (EUV) sources and their applications in challenging scientific and technological problems.

Light in the extreme ultraviolet region of the electromagnetic spectrum covers the 5-50 nm range. Because its wavelength is 100-10 times shorter than visible light it can ‘see’ and ‘write’ smaller patterns in applications such as microscopy and lithography. Furthermore, these wavelengths are well matched to the primary atomic resonances of most elements, making possible many element- and chemically- specific spectroscopies and spectromicroscopies
Our goal is to make EUV light, now mostly limited to a handful of large national facilities, available routinely in a broad variety of laboratory settings, for applications such as high-resolution imaging, spectroscopy, elemental- and bio-microscopy, and nano-fabrication. This picture shows all the different applications the EUV ERC has demonstrated in the last 3 years by combining unique coherent EUV sources with state of the art EUV optics. Through these efforts the EUV ERC is educating a diverse group of students and young scientists in EUV optical technologies who will then go on to play a critical role in maintaining U.S. technological competitiveness.

Award # 0447689 Agency NSF Lead Institution University of IdahoTypeExtramural

Explanatory Notes This project includes work to develop and use nanosensors in aqueous environmental settings. Metrology efforts in the development of these sensors support characterizations of nanomaterials and assessment of their chemical and physical properties.

Abstract The Idaho EPSCoR Research Infrastructure Improvement award will, strengthen shared facilities, build capacity, and enhance research competitiveness in the general area of water quality and uses relating to environmental change - specifically for five integrated research themes: 1) hydrologic processes relating to the global debate on "Old Water" and it's variable chemistry, 2) carbon/water flux at the local soil-vegetation-atmosphere scale, 3) carbon/water export from watersheds and nutrient limitation, 4) fish physiology and genomics, and 5) aqueous environmental nanosensors. A diversity of researchers from the University of Idaho, Idaho State University, and Boise State University will be brought together in the kind of collaborative environment essential for useful and meaningful infrastructure improvement to increase competitiveness for federal research support. A highly instrumented Idaho Experimental Watershed Network will be established; data management and analysis capabilities will be strengthened for the Hydroinformatics Facility of the new Idaho Water Center; and a state-of-the-art regional water quality laboratory will be developed to strengthen collaborations with Idaho and regional Native American tribes as well as national and international researchers. This project will contribute to scientific understanding of environmental geochemical cycles and water quality and potentially to national and global policy formulation.

Undergraduate and graduate students and postdoctoral associates will be involved in research related to the five specified themes and graduate programs will be restructured to further the integration of research and education within the universities. Existing educational and public outreach programs will be strengthened to attract more students to scientific and engineering studies/careers and to increase public awareness of the role of research in higher education and its contribution to the state's economic growth. In particular, current, working relationships with Idaho's Native American Tribes in both research and education will be expanded to other states, engaging the broader Columbia River Inter-Tribal Fish Commission. Outreach efforts to the private sector also include a Phase 0 SBIR program, offering assistance with development and writing of more successful SBIR proposals to different agencies. The project offers strong potential for strengthening and expanding agriculture, aquaculture and related industries in Idaho.

Award # 0335765 Agency NSF Lead Institution Cornell UniversityTypeExtramural

Explanatory Notes This award supports a distributed, multi-faceted, and broadly accessible infrastructure that is being utilized in the development of instrumentation, metrology, and analytical methods to assess EHS aspects of nanomaterials.

Abstract The National Nanotechnology Infrastructure Network (NNIN) is a partnership of 13 institutions (Cornell University, Georgia Institute of Technology, Harvard University, Howard University, North Carolina State University (affiliate), Pennsylvania State University, Stanford University, University of California at Santa Barbara, University of Michigan, University of Minnesota, University of New Mexico, University of Texas at Austin, and University of Washington) that provides multi-faceted, interdisciplinary, and broadly-accessible infrastructure supporting both near-term and long-term needs identified in the National Nanotechnology Initiative. The partnering facilities are open laboratories providing outstanding service to the external user, comprehensive training and staff support, and support of interdisciplinary and emerging areas of research, with openness to new materials, techniques, and applications.

Some of the key scientific, educational, and societal needs supported by the network are:
*Easy implementation of nanotechnology experiments through integration of knowledge and coordination of large numbers of different types of patterning and processing steps, together with the complex tasks of synthesis and assembly at the molecular scale
*Specialized techniques for characterization and metrology at the atomic scale
*Support of advanced and robust modeling and simulation through software and hardware resources with strong technical support
*Technology transfer and the sharing of new techniques through in-person and web-based interactions
*Education and technical training of new users who will be the leaders in nanotechnology
*Public education about the opportunities and challenges of nanotechnology
*Promotion of research in the social sciences so that future developments lead to the greatest possible societal benefits, and societal and ethical studies that focus on research, infrastructure, and the impacts of nanotechnology.
*Nation-wide outreach across age groups and technical interests, with special efforts to reach non-traditional users and under-represented groups.
*Active coordination and knowledge dissemination on safety, environment and health benefits and risks of nanotechnology

Specifically, the network provides:
Infrastructure for Research: The network provides on-site and remote access for users, from academia, small and large industry, and government, to advanced top-down patterning and processing and bottom-up synthesis and self-assembly, comprehensive integration capabilities for multi-step processes, state-of-the-art characterization for hard and soft materials, the development of tools and techniques, and a comprehensive web and computation infrastructure in support of nanotechnology. The network has easy user access that enables a diversity of projects efficiently and at low cost: e.g., molecular-scale electronic contacts; use of functionalized nanotubes; integrated mechanical, electronic, fluidic, and bio-systems; advanced types of microscopy; and a large ensemble of other projects. The network also develops tools in support of future research and nano-manufacturing, including imprint and soft-lithography with applications in electronics, microfluidics, and nanobiotechnology.
Infrastructure for Network Web and Computation: The network's computation and web-based infrastructure provides a centralized resource for organizing and distributing the rapidly-growing knowledge base at the foundation of nanoscience and engineering. It includes training in tools, processing, and synthesis techniques; classes; discussion groups; an open text-book with links to the technical resources and data-bases; and technical support for robust computational tools for design, simulation, and modeling. The web-based infrastructure also comprehensively links our initiatives in education, outreach, and societal and ethical studies in order to provide nation-wide access.
Collaboration and External Interactions: The network employs connections and extensive collaborations with national and industrial laboratories, and with foreign institutions. Through these partnerships and joint meetings and workshops, we share expertise and perspectives, provide specialized training opportunities, coordinate access to unique instrumentation, and transfer newly developed technologies.
Infrastructure for Education, Human Resource Development, Outreach, and Societal and Ethical Studies: Education, human development, outreach, and societal and ethical studies components are thoroughly integrated throughout the network. Our goals are to spread the benefits of nanotechnology to new disciplines, to educate a dynamic workforce in advanced technology, and to become a teaching resource in nanotechnology for people of all ages and educational backgrounds. Network-based education and information tools and comprehensive local hands-on activities towards these goals include: development of a hyper-linked Open Textbook for advanced students, web-based education and virtual research with introductory material for "K-to-gray" distance learning, outreach to 2- and 4-year colleges, a web-based magazine (a mini Scientific American/New Scientist) to interest 6-10 year olds in science, modular teaching packages for nanoscience and for laboratory experimentation in schools, experience-providing programs for undergraduates and teachers, and specialized programs for outreach to women, African-Americans, and Hispanics.
The network also supports an infrastructure and research environment to promote consideration of the societal and ethical consequences of nanotechnology, covering economic, political, educational, environmental, health, safety, legal, security, and cultural implications. A network of scholars in these fields are embedded within the NNIN, with coordinated efforts to foster exchange and discussion, development of web-based resources for research and education, public outreach through the media, and an online archive of technical documents, analysis, transcripts and policy recommendations. The research orientation of the effort explores issues of ethics, communication, workforce change, industrial innovation, and other social implications of nanotechnology

Award # 0610213 Agency NSF Lead Institution Virginia Polytechnic Institute And State UniversityTypeExtramural

Explanatory Notes This is an application of nanomaterials for improving measurements in environmental and occupational safety. Specific work aims to develop higher performance sorbents for gas chromatography through the use of nanostructured thin films.

Abstract Since the 1950s, gas chromatography (GC) has been a common approach for analysis of volatile mixtures in which the components are differentiated in space and time. Conventional GCs tend to be large, fragile, and relatively expensive table-top instruments with high power consumption, but they are known to deliver accurate and selective analysis. The use of MEMS technology for GC development is a promising approach to micro-instruments having lower cost, smaller size, lower power consumption, faster analysis, and greatly increased portability for in-field use. Such systems will make gas chromatography a pervasive method for gas analysis, with applications in homeland security, monitoring food freshness, industrial process control, biomedical diagnostics, and improving environment quality. In GCs, due to low concentration of volatile and semivolatile organic compounds in the environment, a preconcentrator prior to real-time chemical sensor measurement is needed to automatically sample the ambient gas and improve the measurement sensitivity by 10-1000 folds. Miniaturization of preconcentrators using silicon micromachining techniques can overcome the limitations of conventional methods (using a narrow bore metal tubing) by reducing the device size, power consumption, dead volume, and thermal mass. Although achieving promising results, microfabricated preconcentrators still face difficult challenges in achieving a preconcentrator with high adsorbent capacity (>1000), low power consumption (<1W peak-power), and narrow injection plug width (<0.2s). Herein, we will address these challenges by combining and bridging the gap between top-down miniaturized processing and bottom-up self-assembly approaches for the first time to develop miniaturized preconcentrators. The objective of this work is to employ MEMS technology to fabricate preconcentrators having on-chip thermal desorption capability and to utilize nanotechnology to coat the preconcentrator interior surfaces with nano- structured materials such as ionically self-assembled films. Three specific aims are proposed: 1) Fabrication of lowmass (low-power) preconcentrators having integrated heaters and temperature sensors for thermal desportion using high-aspect-ratio silicon etching techniques and a silicon-on-glass process, 2) Deposition of ionic self-assembled multilayers (ISAM) on the preconcentrator walls as adsorbents only a few tens of nanometers thick, and 3) Evaluation of the preconcentrator performance in terms of breakthrough time and volume, concentration factor, and temperature requirements. We envision that the use of nanostructured adsorbents (such as nanoparicles) with high surface to volume ratio ensures that the preconcentrator has sufficient surface area for trapping the sample stream. This reduces the preconcentrator volume and hence decreases the overall mass of the structure. The low-mass preconcentrator allows rapid thermal desorption to generate narrow bands for injection into the GC column.

The broader impacts of this exploratory project will set an outstanding example of how MEMS and Nanotechnology can become highly complementary methodologies to develop low-cost, low power, high-performance devices that impact industries across the globe considering that the worldwide market for GC instruments is estimated to be around $1 billion annually. This research will also advance discovery while promoting teaching and learning at undergraduate and graduate levels. This includes recruiting of graduate students from under-represented groups into a highly interdisciplinary research program, and incorporation of the project results in the courses taught by the PIs in three different departments, namely MEMS: from fabrication to application,Nanotechnology, and Advanced Analytical Chemistry-Separation Science. Additionally, the outcome of this research will be widely disseminated to the engineering and scientific communities in journals and in presentation at multidisciplinary conferences.

Award # Agency DOD (AFOSR) Lead Institution TypeExtramural

Explanatory Notes

Abstract The number of products that contain nanomaterials and the associated risk of exposure to DoD personnel is rapidly increasing. This project will identify factors that influence the generation, dispersion, and deposition of nanomaterials in the workplace. The risk of exposure via inhalation or dermal contact will be quantified for benchmark nanomaterials with well defined physical and chemical characteristics. The effectiveness of personal protective equipment for protecting workers from various release events will be evaluated. A comprehensive guide for workers who handle and use nanomaterials will be prepared to assess, avoid, and abate the adverse health and environmental impacts of nanomaterials

Award # R833328 Agency EPA Lead Institution North Carolina State UniversityTypeExtramural

Explanatory Notes

Abstract Description: The wide applications of manufactured nanomaterials will create enormous potential for human exposure and environmental release. Skin, as the largest organ protecting the body from exogenous toxins and particulates, will be a major portal of entry for nanomaterials. Our preliminary study has shown that fullerene nanoparticles can penetrate deep into the stratum corneum (the primary barrier of the skin) and be modulated by solvents and ion-pairing agents. Currently, there is no method available for quantitative assessment of the skin absorption of the manufactured nanomaterials. Objective: The objective of this project is to establish a structure-permeability relationship for skin absorption of manufactured nanomaterials for safety evaluation and risk assessment. Four dominant physiochemical properties (particle size, surface charge, hydrophobicity and solvent effects) in skin absorption will be studied. Fullerene and its derivatives will be used as model nanomaterials. The absorption and disposition kinetics and dose-response relationships will be measured experimentally for quantitative model development. Approach: The novelty of this project is to study one parameter of interest (e.g., size) while keeping other parameters (e.g., surface charges and hydrophobicity) constant, in contrast to most of the current research focusing on the toxicological effects of the nanomaterials. Three well-developed experimental methods will be used in consideration of throughput, cost and biological complexity. Diffusion experiments will provide in-vitro absorption kinetic information by measuring the nanomaterial flux across the skin. Tape-stripping is designed to provide in-vitro disposition kinetic information of the nanomaterials in the stratum corneum. An isolated perfused porcine skin flap (IPPSF) technique will provide ex-vivo absorption kinetic information that has proven to be effective for human in vivo prediction. Expected Results: The ion-pairing effects, solvent effects, and the impact of particle size and hydrophobicity on skin absorption of nanomaterials will be quantitatively measured to provide three sets of absorption kinetic data: in-vitro absorption, ex-vivo absorption, and in-vitro disposition kinetics. The quantitative data obtained in this project will be used to develop quantitative structure-permeability relationships based on the physiochemical properties of nanomaterials, which will define a general applicable approach for quantitative risk assessment and safety evaluation of manufactured nanomaterials.

Award # 5R33DK066990-03 Agency NIH Lead Institution Northwestern UniversityTypeExtramural

Explanatory Notes This grant will provide data on the implantation of noble metal nanoparticles in the skin.

Abstract NIH estimates approximately 17 million Americans have diabetes mellitus, with approximately 1/10 of these being type 1 and the remaining being classified as type 2, gestational, and other. The disease can be characterized by long-term complications involving eyes, kidneys, nerves, and blood vessels are common but are limited with tight glycemic control characterized by near-normal glycosylated hemoglobin. The goal of the proposed research is to prove that a sensor based on Surface Enhanced Raman Spectroscopy (SERS) can be used to measure glucose levels in vivo with sufficient accuracy for feedback control of insulin delivery. We propose to develop the fiber-optic-based sensor, test the sensor in vitro and in vivo, and acquire sufficient data in vivo to specify the system requirements for use in human trials. Much of the previous work in the field of glucose sensing has focused on minimally invasive means of measuring blood glucose levels; such systems will allow patients to avoid repeated painful finger-pricks but glucose control would still be based on patient-dependant feedback control. Implanted electrochemical sensors have been FDA-cleared since mid-1999, but they must be physician implanted and their use in real-time glucose control has been limited. Our preliminary SERS work, based on noble metal nanoparticles has shown the potential accuracy and precision of a SERS sensor for glucose. We propose that an advanced version of this SERS sensor could be passed through the skin into the subcutaneous space, in a manner similar to the placement of insulin-pump-based catheters, and replaced every 3 days, which is the current norm for insulin-pump catheters. The nanoparticle-based SERS system provides a direct measure of glucose concentration, can monitor glucose levels continuously, is not based upon consumables (e.g. enzymes or substrates), and like all Raman-based analysis is specific to glucose and relatively insensitive to confounding analytes. We envision using these sensors to close the feedback loop by using the continuous measurement of glucose to control insulin levels, thus freeing the patient of repeated measurements, carbohydrate-counting, and insulin-dosing problems

Award # 5R21CA112436-02 Agency NIH Lead Institution Case Western Reserve UniversityTypeExtramural

Explanatory Notes The EHS component of this proposal is to apply a combined theoretical and experimental approach to gain fundamental understanding of the physical principles of targeting by complex polymer nanoparticles by supplying an experimental practitioner with specific recommendations concerning the grafting density, PEG chain length, architecture, nanoparticle size and density of functional groups that ensure effective targeting ofnanoparticles.

Abstract Targeted drug delivery via nanoparticles holds great potential for the successful treatment of many deadly diseases such as cancer. PEG-modification is commonly used to prevent nonspecific interactions of the nanoparticles with blood components and nontarget cells. However the ability to achieve high targeting efficiency at the tumor site remains a significant challenge. There are several reports showing a reduction of transfection efficiency and decreased adhesion of phospholipids, liposomes and polymersomes presumably due to shielding of the targeting functional groups by the PEG chains. PEG chain length, grafting density, nanoparticle size and density of functional groups all were found to strongly influence the success of targeting. To analyze the influence of all these factors experimentally is a very complicated and time-consuming task. Computer simulation may considerably aid in this task by analyzing the influence of multiple parameters, thereby guiding the experimental research. The objective of the present proposal is to apply a combined theoretical and experimental approach to gain fundamental understanding of the physical principles of targeting by complex polymer nanoparticles. The central hypothesis is that the ligand valence and the structure of the corona of nanoparticles are the key factors defining the efficiency of site-specific targeting. To test this hypothesis we will carry out research designed to fulfill the following aims: 1) Structural design of a nanoparticle for effective targeting for each of the following concepts: a) using multivalent ligands and b) using short non-functional and long functionalized PEG chains in nanoparticle corona. 2) Design and experimental testing of the optimal PCL-PEG-cRGD nanoparticles for doxorubicih (DOX) delivery. Nanoparticle designs found to be optimal in simulations will be tested experimentally using in vitro binding study and cell uptake study in integrin-expressing and non-expressing melanoma cells. Successful execution of this research will supply an experimental practitioner with specific recommendations concerning the grafting density, PEG chain length, architecture, nanoparticle size and density of functional groups to ensure effective targeting of drug-containing nanoparticles. Because of the exploratory nature of the research some new approaches to drug targeting may be discovered. This knowledge will greatly facilitate the development and implementation of a new generation of drug delivery vehicles for cancer therapy

Award # 5R21CA114143-02 Agency NIH Lead Institution University of WashingtonTypeExtramural

Explanatory Notes The goal of this research is to harness forces generated by actin polymerization to propel nanoparticles within the interstitial space by energy-mediated, cell-to-cell transfer, thus resulting in more efficient nanoparticle penetration. The EHS-relevant component of this research will demonstrate modifications that produce efficient delivery systems.

Abstract New technologies for molecular analysis of cancer identify patterns of genetic and protein expression changes that have occurred in tumorigenic cells. Application of these tools for in vivo analysis is critical for a complete understanding of metastatic cancer; sadly, such studies have been limited by the lack of effective methods for delivery to metastases. Nanoparticle formulations of these agents offer in vivo protection and concentrated tumor delivery and are therefore promising delivery entities. However, a major limitation of nanoparticles for tumor delivery is restricted interstitial transport. Here, we propose to harness forces generated by actin polymerization to propel nanoparticles within the interstitial space by energy-mediated, cell-to-cell transfer, thus resulting in more efficient nanoparticle penetration. This goal can be achieved by realizing the following aims: (i) modifying nanoparticles with ActA, a bacterial protein that initiates actin polymerization resulting in propulsive forces, and optimizing formulations for motility in cytoplasmic extract, (ii) achieving actin-mediated, cell-to-cell transfer of nanoparticles in cultured monolayer cells, and (iii) demonstrating improved nanoparticle penetration in three-dimensional spheroid cultures. Efficient delivery systems are crucial for both research and clinical applications; thus, successful completion of this project would result in a major step toward realizing the full potential of molecular analysis, detection, and treatment of cancer

Award # 5R01CA101850-04 Agency NIH Lead Institution University of UtahTypeExtramural

Explanatory Notes The primary purpose of this research is to engineer functional polymeric micelles which target solid tumors in acidic extracellular fluid and utilize acidic endosome to treat sensitive and multidrug resistant tumors by designing biodegradable polymers sensitive to tumor acidity and to engineer polymeric micelles with or without targeting moiety that can truly recognize tumor pile or endosomal pH for triggered release, while keeping a minimal release rate during circulation.

Abstract The primary function of this application is to engineer functional polymeric micelles which target solid tumors in acidic extracellular fluid and utilize acidic endosome to treat sensitive and multidrug resistant (MDR) tumors. It is estimated that more than 80% of measured tumor extracellular pH (pile) are below 7.2. For intracellular pH of tumor cells, parenteral drug sensitive cells are characterized to have rather acidic, diffuse cytosolic pH profile; however MDR cells develop more acidic organelles (recycling endosome, lysosome and trans-Golgi network) than cytosol and necleoplasmic pH. Our preliminary results demonstrate that the polymeric micelles composed of poly(L-histidine)/PEG and PLLA/PEG enhanced the release rate of a loaded model anticancer drug (doxorubicin (DOX) in this study) by physical destabilization of the micelle core at pile, resulting in higher cytotoxicity at lower pH. In addition, the micelles, conjugated with folate and destabilized at pH 6.8, showed great efficacy for sensitive and MDR cells after folate receptor-mediated endocytosis. Therefore it is hypothesized that triggered release of DOX from the intelligent polymeric micelles at tumor pile is a more effective modality in cancer chemotherapy, proving higher local concentration at tumor sites (targeted high-dose chemotherapy), while a minimal release during circulation occurs. The micelle destabilization may help further accumulation of the micelles by reducing the physical barriers in the interstitial space. Another hypothesis is that after receptor-mediated endocytosis, simultaneous triggered release in early endosomes (approximately pH 6) and endosomal disruption will provide high concentrations of the drug in cytosol and nucleus. This will be effective not only for sensitive and but for MDR cells where the drug diffusivity the plasma membrane is compromised and the pumping activities of Pgp and MRP. This approach will be especially useful for weakly basic drugs of which partitioning between cytosol and subcellular organelles is greatly influenced by pH gradient (sequestration). The goals of this research are 1) to design biodegradable polymers sensitive to tumor acidity and to engineer polymeric micelles with or without targeting moiety that can truly recognize tumor pile or endosomal pH for triggered release, while keeping a minimal release rate during circulation and 2) to assess the proposed hypotheses for improved chemotherapy

Award # 5U54CA119343-02 Agency NIH Lead Institution University of North Carolina-Chapel HillTypeExtramural

Explanatory Notes This project will bring together nano-particle engineering with the understanding/treatment of cancer for the delivery of therapeutic, detection and imaging agents for the diagnosis and treatment of cancer by fabricating "smart" functional particles for studies and evaluations. This will allow the fabrication of nano-biomaterials to accelerate translational understanding, detection and treatment of cancer.

Abstract The Carolina Center of Cancer Nanotechnology Excellence (C-CCNE) will bring together recent pioneering breakthroughs at UNC in nano-particle engineering with the world class excellence in the understanding/treatment of cancer in the Lineberger Cancer Center for the delivery of therapeutic, detection and imaging agents for the diagnosis and treatment of cancer. Physical scientists at UNC have pioneered a recent breakthrough in the materials useful for imprint lithography-an emerging technology adapted from the microelectronics industry-that enables an extremely versatile and flexible method for the direct fabrication and harvesting of monodisperse, shape-specific nano-biomaterials. The method, referred to as Particle Replication In Non-wetting Templates, or PRINT, allows for the fabrication of monodisperse particles with simultaneous control over structure (i.e. shape, size, composition) and function (i.e. cargo, surface structure). Unlike other particle fabrication techniques, PRINT is delicate and general enough to be compatible with a variety of important next-generation cancer therapeutic, detection and imaging agents, including various cargos (e.g. DNA, proteins, chemotherapy drugs, biosensor dyes, radio-markers, contrast agents), targeting ligands (e.g. antibodies, cell targeting peptides) and functional matrix materials (e.g. bioabsorbable polymers, stimuli responsive matrices, etc). Within the C-CCNE, researchers in the Lineberger Cancer Center and the Schools of Medicine and Pharmacy will use specially designed PRINT particles to reach new understandings and therapies in cancer prevention, detection, diagnosis and treatment. The C-CCNE will bring together researchers and clinicians not only at UNC but nationally and internationally to meet the Challenge Goal of eliminating the suffering and death from cancer by 2015. This will be accomplished by establishing a national NCI asset we refer to as the PARTICLE FOUNDRY. The PARTICLE FOUNDRY will be one of the outcomes of this project within the Center of Cancer Nanotechnology Excellence at UNC. The PARTICLE FOUNDRY will be the portal for researchers around the world to gain access to UNC's breakthrough PRINT technology for the fabrication of "smart" functional particles for their studies and evaluations. As such, PRINT is a significant scientific and technological breakthrough which will allow the fabrication of heretofore inaccessible populations of nano-biomaterials which are poised to revolutionize and accelerate our translational understanding, detection and treatment of cancer

Award # 5U54CA119335-02 Agency NIH Lead Institution University of California-San DiegoTypeExtramural

Explanatory Notes This project seeks to develop a multi-functionalplatform that delivers payloads of both nanosensors and therapeutics directly to a tumor.The EHS-relevant component project focuses on the production of tumor-targeted, non-toxic nanoparticles

Abstract The Center of Excellence of NANOtechnology for Treatment, Understanding, and Monitoring of Cancer (NANO-TUMOR) has been established to perfect a practical nanotechnology base to diagnose, treat, and monitor cancers. Our goal is the development, of a multi-functional "smart mothership" platform that will: (1) evade the reticuloendothelial system and immune system, while attaching specifically to the tumor and its vasculature; (2) assemble a multi-functional complex at the tumor site; (3) deliver payloads of both nanosensors and therapeutics that are activated in situ. The NANO-TUMOR Center has brought together a team of investigators from the University of California, San Diego, Santa Barbara, Irvine, and Riverside campuses, the Burnham Institute and NanoBioNexus. The NANO-TUMOR Center research program is comprised of six interacting projects, each utilizing newly developed and unique technologies that when integrated together will lead to the realization of our goal. The six projects focus on (1) the production of tumor-targeted, non-toxic nanoparticles, (2) the development of a nanoporous micro-platform carrying nanosensors, imaging and therapeutic agents, (3) the creation of tumor-directed amplification systems for sensing and drug delivery, (4) the construction of devices for tumor molecule purification and characterization at a nanoscale level, (5) the application of new computational methods for longitudinal monitoring and analyses of tumor progression and response to therapy, and (6) the in situ assembly and delivery of targeted therapeutics with a smart nanotechnology platform. The Center has been organized using a novel, engineering-type decision and workflow model: Multiple approaches have been proposed to address each project, with go/no go decisions made based on achievement of milestones. The NANOTUMOR project leaders have previously founded more than 20 successful biotechnology companies, with a total market value of over two billion dollars, evidence of their ability to move discoveries to useful applications in the marketplace. The intent of the NANO-TUMOR Center to develop clinically useful platforms is also demonstrated by the strategic role that clinicians from the UCSD Cancer Center, and participants from major corporations (General Electric, Honeywell, Nanogen, Enterprise) will play in evaluating the research

Award # 5U54CA119338-02 Agency NIH Lead Institution Emory UniversityTypeExtramural

Explanatory Notes This EHS component of this research integrates nanotechnology with cancer biomolecular signatures and will provide data on the interaction of nanomaterials with cells.

Abstract This application proposes a cross-disciplinary Center of Cancer Nanotechnology Excellence (CCNE) at Emory University and Georgia Tech that will integrate nanotechnology with cancer biomolecular signatures (biomarkers) for personalized and predictive oncology. The overarching scientific focus is to accelerate the development of bioconjugated nanoparticles for cancer molecular imaging, molecular profiling, and personalized therapy. The proposed research will further develop new nanoparticle reagents for co-targeting signal transduction pathways and its microenvironments that are involved in bone metastasis. This research is broadly applicable to many types of malignant tumors such as lung cancer, colorectal carcinoma, ovarian cancer, brain tumors, and lymphomas; but proof of concept research is focused on human prostate and breast cancers and their clinically aggressive phenotypes - including interrogations of cancer cells and patient tissue biopsies. A compelling reason for this focus is that breast and prostate cancers represent a number of compelling challenges and opportunities in human oncology such as high incidence and mortality, evidence that targeted therapies can improve survival in these cancers, and our linkages to NCI Specialized Programs of Research Excellence (SPORE) in both prostate and breast cancers. The CCNE has 6 "synergistic projects" with crossdisciplinary teams, each with expertise in nanotechnology, bioengineering, clinical oncology, and basic cancer biology. Project #1 will develop quantum dots and targeted nanoparticles for cancer molecular imaging (Nie). Project #2 will develop novel molecular beacons and activatable nanoprobes for gene expression studies of single cancer cells (Bao). Project #3 will optimize and translate "nanotyping" multicolor sets of antibody linked quantum dots to multiplex cancer biomarkers that can predict clinical outcomes and susceptibility to signal transduction inhibitors in medical oncology (Simons/O'Regan). Project #4 will develop surface-enhanced Raman spectroscopic (SERS) nanotags and atomic nanoclusters for molecular profiling in cancer pathology (Natan/Young). Project #5 will develop nanoparticle anti-cancer therapeutics using a new class of self-assembled and biodegradable nanoparticles (Shin). Project #6 will focus on basic cancer metastasis biology and creation of new reagents via bioconjugated nanoparticles to target metastatic cancer cell clones and their bone stromal microenvironments (Chung). These projects are supported by 5 core functions: nanomaterials synthesis and fabrication (Core 1 - Z. Wang); biocomputing and bioinformatics (Core 2 - M. Wang); tissue specimens and animal tumor models (Core 3 - Datta); onconanotechnology education and outreach (Core 4 - Simons); and center administration, biostatistics support, technology assessment, and commercialization (Core 5 - Nie/Simons/Murdock). This CCNE is strengthened by collaborations with three NCI CCCs, investment from the Georgia Research Alliance, Georgia Cancer Coalition, and industrial partners. The CCNE is embedded in the Winship Cancer Institute, a new integrated 280K sq ft cancer research and care building, and has a special constellation of US partners, the American Cancer Society and the Centers for Disease Control, for accelerating the discovery and clinical translation of nanotechnology to reducing the burden of human cancer

Award # 5U54CA119341-02 Agency NIH Lead Institution Northwestern UniversityTypeExtramural

Explanatory Notes This project involves the use of nanomaterials in living systems and the evaluation of the biocompatibility and biodistribution of materials in the body and within cells.

Abstract The potential impact of nanotechnology is well recognized, and significant advances in the medical field are expected to be realized first. It is possible that nanotechnology will be the fundamental driver of advances in oncology and cancer research leading to near-term benefits for patients, and yet formidable challenges must be met. Northwestern University proposes to meet these challenges through the establishment of a Nanomaterials for Cancer Diagnostics and Therapeutics Center for Cancer Nanotechnology Excellence (CCNE). To be led by the Principal Investigator, Chad A. Mirkin, the proposed CCNE represents the development of a strong integrated partnership between the NU Robert H. Lurie Comprehensive Cancer Center (RHLCCC), and the NU International Institute for Nanotechnology (IIN). The RHLCCC is an NCI designated, comprehensive, University-based, matrix cancer center conducting a broad range of multidisciplinary basic, clinical, and population science research with over $116 million dollars in annual extramural funding. The NU International Institute for Nanotechnology (IIN) is an umbrella organization which unites all of the nanotechnology research and educational programs at NU, and encourages and supports collaborations with the Center for Nanoscale Materials (CNM) at Argonne National Laboratory. Investigators in the NU IIN currently are supported by more than $130 million in extramural funding. Building upon the significant advances in cancer research and in nanotechnology - particularly in the detection arena - obtained at NU, and operating within the framework of a single university will permit this CCNE to optimize the intensive level of integration and collaboration required to create an accelerated pathway-from conception to clinical trial-for development of nanomaterials and nanodevices to overcome cancer. Other academic collaborators include the University of Chicago, the University of Illinois/Urbana-Champaign, and Yonsei University, South Korea

Award # 5U54CA119349-02 Agency NIH Lead Institution Massachusetts Institute of TechnologyTypeExtramural

Explanatory Notes This grant has a Toxicity Core and a Mouse Models Core that will provide in vivo information about nanomaterials behavior in model systems.

Abstract The overall goal of this U54 application is to create and support a highly multidisciplinary team of chemists, biologists, engineers and physicians to develop and rapidly translate new nanotechnologies to better diagnose and treat cancer. The current team includes investigators from Massachusetts Institute of Technology (MIT), Harvard Faculty of Arts and Sciences (FAS), Harvard Medical School (HMS), Massachusetts General Hospital (MGH), and Brigham and Women's Hospital (BWH). Specific applications of nanotechnology in this application include targeted therapies, diagnostics, noninvasive imaging, and molecular sensing.Project 1 (Langer, Farokhazad) is developing novel nanoparticles for cancer targeting using prostate cancer as a model. Project 2 (Sharp, Bhatia) is developing new siRNA delivery and targeting strategies for use in treatment of cancer (lung and brain cancers will serve as models). Project 3 (Weissleder, Josephson) is developing clinically viable, next-generation magnetic nanoparticles for targeted multimodal imaging of cancer. Project 4 (Cima) is creating unique miniaturized MEMS-based devices for molecular sensing. Project 5 (Belcher, Bawendi) is developing and applying novel semiconductor nanocrystals for biomedical sensing in the context of cancer. In the first year, up to eight pilot projects are being developed as a means of attracting new investigators to the consortium, to stimulate creative, high-impact research, to rapidly test new nanomaterials, and to fund collaborative work. The Projects and Pilot Projects are supported by a Toxicity Core and a Mouse Models Core. It is anticipated that this research in nanotechnology will significantly advance medical science and treatment of cancer.

Award # 5U54CA119342-02 Agency NIH Lead Institution Washington UniversityTypeExtramural

Explanatory Notes The goal of this project is to employ nanoparticles for targeted delivery of chemotherapeutic agents and for imaging.The EHS-relevant component is the mechanistic understanding of nanoparticle uptake and cell interaction.

Abstract Over the last thirty years many noteworthy advances in the early detection of breast, colon, and prostate cancer have improved treatment and, in some instances, improved outcomes. Yet during this timeframe, the incidence of cancer continued to increase, morbidity from treatments (i.e., surgery, radiation and chemotherapy remained unacceptable, and survival only improved marginally. The subject of this proposal is the application of a novel paramagnetic site-targeted contrast "platform technology" for sensitive and specific imaging of molecular epitopes expressed on tumor neovasculature alone and in combination with the local delivery of chemotherapeutic agents to these sites. ava3-integrin nanoparticles effectively target solid animal tumors and human xenografts to provide marked MR T1-weighted contrast and potent anti-tumor therapy. Unfortunately, no single vascular biosignature is ubiquitous across all solid tumors. Therefore, tumors must be noninvasively interrogated against a broader panel of targeted agents in order to individualize therapy with the appropriate single or combination of ligand directed nanoparticles. This program, we will expand utility of this successful nanotechnology platform to additional early biosignatures that may be targeted alone or simultaneously for the most effective diagnostic and chemotherapeutic response. In parallel, we will develop noninvasive imaging software and hardware to exploit the unique opportunity presented by perfluorocarbon nanoparticles for 19F MR spectroscopy, 19F imaging, and 1H/19F hybrid imaging. The use of 19F spectroscopy and imaging will not only add quantitative and spectral dimensionality to targeted nanoparticle applications, but 19F imaging can also eliminate the need for baseline digital subtraction of images, which is time-consuming and prone to error. Interleaved 1H/19F hybrid imaging will minimize motion artifacts, eliminate image registration issues, confirm contrast identification and shorten patient scanning times

Award # 1U54CA119367-01 Agency NIH Lead Institution Stanford UniversityTypeExtramural

Explanatory Notes The goal of this project is to employ nanoparticles for imaging. The EHS-relevant component is the mechanistic understanding of nanoparticle uptake and cell interaction.

Abstract This Center for Cancer Nanotechnology Excellence focused on therapy response (CCNE-TR) brings together scientists and physicians from Stanford University, University of California Los Angeles (UCLA), Cedars Sinai Medical Center, Fredrick Hutchinson Cancer Center, University of Texas at Austin, Intel Corporation, and General Electric Global Research in a novel proposal to utilize nanotechnology for the benefit of cancer patient management. This research proposal is centered around our vision that ex vivo diagnostics used in conjunction with in vivo diagnostics can markedly impact future cancer patient management. Furthermore, we believe that nanotechnoloqy can significantly advance both ex vivo diagnostics through proteomic nanosensors and in vivo diagnostics through nanoparticles for molecular imaging. The cancer-related biochemical pathways targeted will be the Her-kinase axis with prostate cancer as the initial focus, and CD20/c-myc with lymphoma as the second initial major focus. We have assembled a highly interdisciplinary team of scientists from the fields of chemistry, materials science and engineering, molecular imaging, oncology, cancer biology, protein engineering, biostatistics, and mathematical modeling in order to accomplish our goals. We highly leverage resources at the Stanford Bio-X Program, National Nanotechnology Infrastructure Network, the California Nanosystems Institute, the Stanford/UCLA/Fred Hutchinson Cancer Centers, as well as significant resources with our two primary industrial partners (GE and Intel). We also have direct links to a UCLA Prostate SPORE, the ICMIC P50 and ICBP P20 at Stanford as well as several other NCI sponsored efforts. Furthermore, we have many methods for outreach and dissemination including the Prostate Cancer Foundation (formerly named CaPCure) as well as the Canary Foundation. Projects will focus on the use of magnetonanotechnology and nanotube/nanowire technology for ex vivo protein detection; the use of Raman sensors for protein phosphorylation detection; methods to determine protein profiles on the cell surface, the secretome, and serum from mouse models and humans; the use of biologically targeted quantum dots for molecular imaging of living subjects; and mouse models for integrating ex vivo tissue/serum protein patterns and in vivo molecular imaging to predict response to anti-cancer therapy. Cores provide a nanocharacterization laboratory resource; service for fabricated nanostructures; and an informatics and biostatistics resource. Together, these highly interactive and cohesive programs will produce breakthroughs towards our vision of developing and validating nanotechnology for anti-cancer therapy response

Award # 5R01CA119409-02 Agency NIH Lead Institution University of Michigan-Ann ArborTypeExtramural

Explanatory Notes The goal of this project is to employ dendrimer modules linked by oligonucleotides for targeted delivery of chemotherapeutic agents and for imaging.The EHS-relevant component is the mechanistic understanding of nanoparticle uptake and cell interaction.

Abstract Targeted dendrimer-based nanodevices have shown excellent promise in both in vitro cell culture and in vivo animal studies as cancer therapeutics. However, each device must be custom synthesized for a particular set of targeting molecules, imaging agents, and desired therapeutics. We propose a unique solution to this limitation by developing single function dendrimer modules linked by complementary oligonucleotides. This allows targeting, imaging, and therapeutic dendrimers to be combined into multifunctional therapeutics simply by heating mixtures of these agents above the annealing temperature of the oligonucleotide duplex. The project will consist of five specific aims. Specific Aim 1 will involve the design and synthesis of complementary oligonucleotides conjugated to poly(amido)amine dendrimers or dendrons to achieve the desired structural topologies. The ability to construct complex devices will be assessed using well-defined targeting molecules (folic acid, her2 antibodies and RGD peptides), drugs (methotrexate, Taxol, cis-platin and doxorubicin), imaging agents (Gadolinium chelators and fluorescent dyes). Specific Aim 2 will characterize the self-assembled nanodevices using techniques including PAGE, HPLC, CE, Mass Spectroscopy, NMR, AFM, and NSOM. Specific Aim 3 involves testing the DNA-linked nanodevices for binding and internalization in vitro; the avidity and specificity of binding will be examined using CD, differential calorimetry and Biacore analyses. Devices carrying therapeutics will be tested for effectiveness at inducing cell death, and all devices will also be tested for inherent cytotoxicity. Specific Aim 4 employs animal models to assess the effectiveness of the dendrimer linked therapeutics to treat tumors in vivo. In addition, the biodistribution of the therapeutics will be assessed using radiolabeled material and a novel fiber optic probe that uses two-photon excitation with femto-second pulses. Finally, under Specific Aim 5 we will work with the NCI nanoparticle characterization lab in Frederick to make the materials developed in this program available to other investigators. This platform has the potential to revolutionize cancer therapeutics and facilitate "personalized medicine." Lay description: We are designing a method and the tools for developing targeted cancer drugs that can be tailored to the needs of an individual patient. The physician can select various components and the components are then linked together like "tinker toys" to make a personalized medicine. This medicine would selectively target only the cancer, thereby avoiding the nausea, hair loss and illness caused by regular cancer chemotherapy

Award # 5R01CA119617-02 Agency NIH Lead Institution Northeastern UniversityTypeExtramural

Explanatory Notes The study seeks to overcome multi-drug resistance in chemotherapy via a multifunctional approach to optimize delivery of pro-apoptotic drugs. The EHS-relevant component of the study will develop, characterize, and optimize long-circulating, biodegradable polymeric nanocarriers and evaluate the uptake, distribution, and intracellular concentrations of carried drugs.

Abstract The development of multi-drug resistance (MDR) is a major cause of failure in chemotherapeutic management of cancer. In breast cancer, for instance, more than 50% of the patients relapse due to acquired resistance to standard chemotherapy regimens. Novel strategies to overcome MDR in a clinically meaningful way that does not expose the patients to significant toxicity are urgently needed. In the proposed study, our strategy to overcome MDR in vivo relies on a multifunctional approach to optimize delivery of pro-apoptotic drugs to the tumor mass, increase the intracellular drug concentrations, and reverse cellular resistance by modulating ceramide levels. The preliminary studies show that we can prepare tumor-targeted biodegradable polymer-based engineered nanocarriers (PENS) for encapsulation of hydrophobic pro-apoptotic drugs, like paclitaxel. We have also found that increasing intracellular ceramide concentrations by delivery from exogenous source or inhibiting the metabolism results in significant enhancement of cytotoxicity in sensitive and resistant tumor cells. C6-ceramide and tamoxifen, a potent inhibitor of ceramide metabolism, were co-administered with paclitaxel for synergistic activity in tumor cells and in vivo. Based on these preliminary findings, we are confident that PENS, developed using engineering design criteria, can be made to efficiently deliver multiple therapeutic agents to the tumor mass. The specific aims of the proposal are: (1) to develop, characterize, and optimize long-circulating, biodegradable polymeric nanocarriers with encapsulated paclitaxel, ceramide, and tamoxifen, either alone or in combination, (2) to evaluate the uptake, distribution, intracellular concentrations of paclitaxel, ceramide, and tamoxifen, cytotoxicity, and apoptotic activity in culture of sensitive and resistant tumor cells, (3) to examine the biodistribution and pharmacokinetic profiles of drugs administered in the control and nanocarrier formulations in sensitive and resistant xenograft tumor models established in nude mice, (4) to determine the antitumor efficacy of single and combination therapy in PENS in sensitive and resistant xenograft models, and (5) use mathematical modeling to improve the design of nanocarriers for tumor-targeted delivery of single and combination drug therapy. The results of this study would be extremely valuable in the treatment of refractory tumors using a multifunctional nanotherapeutic approach that efficiently delivers the drug and can overcome cellular resistance. The multimodal nanocarrier strategy proposed here would provide a translatable approach to overcome MDR in cancer patients.

Award # 5R01CA119387-02 Agency NIH Lead Institution University of Texas-MD AndersonTypeExtramural

Explanatory Notes The goal of this project is to develop novel nanoparticles for molecular optical imaging applications for human cancers by establishing the effect of particle characteristics on the pharmacokinetics, biodistribution, clearance, extravasation, and intratumoral distribution of the nanoparticles.

Abstract Near-infrared fluorescence (NIRF)-based optical imaging of human cancers has several advantages over standard imaging techniques in that it is extremely sensitive, inexpensive, and robust; involves no harmful radiation; and allows real-time visualization. The development of well-validated NIRF imaging probes may lead to this new modality becoming clinically viable for molecular imaging. Recent advances in nanotechnology are likely to substantially accelerate the discovery of new NIRF imaging agents that can not only provide increased signal intensity, but also target or "report" both the presence and the biologic activity of tumor-specific biomarkers. In this application, U. T. M. D Anderson Cancer Center, a leading institution in cancer research and patient care, and Eastman Kodak Co., a world leader in the fabrication of optical dyes and nanoparticles, will team to develop novel nanoparticles for molecular optical imaging applications. Our goals are to systematically investigate the in vivo pharmacologic properties of nanoparticles derived from Kodak's platform technology, and to design and develop nanoparticles that use targeting, enzyme activation, or a combination of both features to achieve significant improvements in the sensitivity and specificity of cancer detection. Our specific aims are 1) to synthesize and characterize polymer-shelled silica nanoparticles and cross-linked PEG nanoparticles suitable for NIRF imaging; 2) to establish the effect of particle characteristics on the pharmacokinetics, biodistribution, clearance, extravasation, and intratumoral distribution of NIRF nanoparticles; 3) to establish the stability and signal intensity of NIRF nanoparticles in vivo and the specificity of their retention in tumors; 4) to construct NIRF nanoparticles targeted to angiogenic blood vessels and to tumor cell-associated surface receptors; and 5) to develop smart, activatable NIRF nanoparticles and to combine homing ligand and molecular beacon designs in a single nanoparticulate system. By using a combination of nuclear and optical imaging, autoradiography, and fluorescence microscopy, we expect to provide detailed insights into the pharmacologic properties of NIRF nanoparticles, with the ultimate goal of obtaining nanoparticles that are effective and practical for molecular optical imaging of human cancers

Award # 5R01CA097465-03 Agency NIH Lead Institution University of UtahTypeExtramural

Explanatory Notes The goal of this project is to employ gadolinium nanoparticles for targeted delivery of chemotherapeutic agents and for imaging.The EHS-relevant component is the mechanistic understanding of nanoparticle biodistribution and clearance.

Abstract The objectives of this research are to develop safe, effective targeted polymer-gadolinium chelate conjugates as contrast agents for magnetic resonance diagnostic cancer imaging. Cancer is one of the leading causes of human death in the United States. The accurate diagnosis and subsequent optimal treatment of cancer at its earliest stage is crucial to save the lives of cancer patients. Magnetic resonance imaging (MRI) is a non-invasive clinical diagnostic technique, but lacks of functional contrast agents for earlier tumor detection and characterization. The novel targeted polymeric contrast agents will increase the sensitivity and accuracy of the detection of small cancers and precancerous tissues with MRI. The specific aims are to design, synthesize and characterize poly(L-glutamic acid)-gadolinium (111) chelate conjugates and monoclonal antibody Fab' fragment targeted poly(L-glutamic acid)-Gd(lll) chelate conjugates as MRI contrast agents with minimal long-term Gd(lll) tissue accumulation; to evaluate the physicochemical and biological properties, including molecular weight distribution, relaxivity, Gd chelate release, in vivo clearance, biodistribution, neoplastic targeting, pharmacokinetics and safety, of the targeted and nontargeted conjugates; to assess the efficacy of the targeted and nontargeted conjugates on tumor contrast enhancement in MR imaging. The biocompatible poly(L-glutamic acid) will be used as a carrier and Gd(III)DOTA will be conjugated to the polymer via cleavable spacers, which will be readily cleaved to release the Gd chelate from the polymer and to facilitate its clearance from the body after the MRI exam. Monoclonal antibody Fab' fragments against the molecular markers expressed in neoplastic tissues will be incorporated into the polymer conjugates to achieve tumor specific contrast enhancement in MRI. The targeted contrast agents will be able to bind to the molecular targets and deliver a sufficient amount of paramagnetic chelates in target tissues for the imaging of small cancers. The structural, physicochemical and biological properties of the conjugates will be further optimized to develop safe, efficacious contrast agents for more accurate cancer detection and staging with MRI. The long-term goal of this project is to develop safe, effective, functional contrast agents for clinical MR cancer imaging

Award # 1R21CA118778-01 Agency NIH Lead Institution Rice UniversityTypeExtramural

Explanatory Notes This grant will evaluate biodistribution, biocompatibility and tumor ablation efficacy of these NIR-absorbing nanoparticles.

Abstract For over fifty years, cancer has remained the second leading cause of death in the United States, accounting for over 25% of the deaths in the population. More than one million cases are diagnosed each year, resulting in over 500,000 deaths (American Cancer Society, 2001). Nanotechnology may offer new options for both diagnosis and treatment of cancer. A new nanoparticle-based approach to cancer therapy has been under investigation in our laboratory. Near infrared-absorbing nanoparticles are injected intravenously and allowed to accumulate in the tumor, due to the leaky vasculature and/or targeting, followed by illumination of the animal with near infrared (NIR) light. NIR light is not appreciably absorbed by tissue components, allowing deep penetration without damage to normal tissues. Using a class of NIR-absorbing nanoparticles called gold-silica nanoshells, we have demonstrated complete tumor ablation and long term survival of animals without tumor regrowth. We have recently begun in vitro studies with two other classes of NIR- absorbing nanoparticles - gold-gold sulfide nanoshells and gold nanorods. In vitro, we have been able to achieve much more rapid heating with these two newer types of nanoparticles due to their higher absorption and lower scattering (order of heating, nanorods, gold-gold sulfide nanoshells, gold-silica nanoshells), and thus believe that they have the potential to be more effective at lower doses in cancer therapy than gold- silica nanoshells. However, the sizes of these three classes of nanoparticles varies widely. Gold-silica nanoshells are approximately 100 nm in diameter, gold-gold sulfide nanoshells approximately 50 nm, and gold nanorods 20 nm. Thus, biodistribution of the particles will be quite different and may drastically affect therapeutic efficacy. We propose to evaluate biodistribution, biocompatibility and tumor ablation efficacy of these NIR-absorbing nanoparticles. The optimal particle formulation will be further examined in a model of medulloblastoma

Award # 1R41CA121453-01 Agency NIH Lead Institution Synergene Therapeutics, Inc.TypeExtramural

Explanatory Notes The EHS component will better characterizenano-immuno-liposomesin animal models, determine optimized dose, optimal time to imaging, and perform toxicity studies in mice.

Abstract The low rate of cure of pancreatic carcinoma is an important health problem. The American Cancer Society estimates that 32,000 Americans will be diagnosed with carcinoma of the pancreas in 2005, and 31,800 will die. Pancreatic cancer is the fourth leading cause of cancer death in the US. Early detection appears currently to be the only way of improving the high mortality rate, but is quite difficult because of the retroperitoneal location of the pancreas and the lack of symptoms in early disease. We are proposing to further develop a MR tumor targeting imaging agent with a high affinity for entering carcinoma cells. Current imaging methods for the pancreas include ultrasound, CT, MRI, and PET. Each of these is moderately to very good for imaging the pancreas and for staging pancreatic neoplasm, but each relies on indirect signs of carcinoma based on the identification of a mass, lesion vascularity, or its glucose utilization. These features are also present in the main mimicker of pancreatic carcinoma; benign masses from chronic pancreatitis. Anti-transferrin Receptor scFv-antibody fragment (TfRscFv) immunoliposome complex is a nano-construct (<300 nm) for delivery of gene therapy to tumors. It has been shown to target human pancreatic carcinoma cell lines in vivo when implanted as xenografts in athymic nude mice. By placing gadopentetate dimeglumine ("gad-d") into the immunoliposome nanocomplex, new and unique capabilities result in the delivery of gad-d directly into tumor cells which express high levels of the transferrin receptor (TfR). Most normal cells have little or no uptake of this agent. Most human pancreatic carcinoma cell lines overexpress the transferrin receptor. The TfRscFv-immunoiiposome gad-d nanocomplex (scL-gad-d) is biodegradable and the residual components are excreted by the kidney (gad-d) or secreted into the biliary system (liposome). The scL-gad-d demonstrates a high sensitivity for tumor targeting and high specificity having very low uptake into cells in the normal tissues of mice. In our preliminary work with this novel tumor targeting contrast media, we see very high tumor to background uptake in pancreatic carcinoma bearing animals (3 times that of standard gad-d). In this Phase 1 STTR project we will better characterize this agent in animal models, determine optimized dose, optimal time to imaging, and perform toxicity studies in mice in preparation for a future Phase 2 STTR submission to perform a Clinical Phase 1 dose escalation toxicity study

Award # 5R01DK064850-04 Agency NIH Lead Institution Massachusetts General HospitalTypeExtramural

Explanatory Notes The goal of this project is to use iron oxide nanoparticles for enhanced MR imaging. The EHS-relevant component of this research is the mechanistic understanding of cell uptake of nanoparticles.

Abstract The overall goal of this proposal is to develop a non-invasive method to detect the infiltration of CD8+ T-cells responsible for beta-cell destruction in pancreatic islets in Type 1 Diabetes (IDDM) by MR imaging. In vivo imaging of immune cells infiltration in real time would have significant impact in managing clinical IDDM, pancreas and/or islet cell transplantation and the understanding of the pathogenesis of IDDM. Unfortunately, such non-invasive techniques are currently not available. Based on our prior experience in cell labeling with crosslinked, superparamagnetic, monocrystalline iron oxide nanoparticles, we synthesized a novel MR imaging probe with high affinity ligands to T cell autoantigenic markers that allowed for high-efficiency intracellular magnetic labeling of the cells with subsequent detection by MRI. This probe specifically binds to highly diabetogenic TCR on CD8+ T cells from NOD transgenic mice, but does not recognize CD8+ T cells from healthy NOD mice. Furthermore, in our initial imaging experiments in vivo we were able to see gradual accumulation of labeled CD8+ T cells in mouse pancreas after adoptive transfer. In the current proposal we are going to conduct imaging experiments using this probe to answer highly relevant biological questions regarding the mechanisms that drive the recruitment, activation and differentiation of autoreactive CD8+ T cells. Specifically, we propose to image in vivo tracking of CD8+ T cell recruitment to the islets and beta-cell mass loss depending on the avidity of recruited killer cells. The proposed studies are a logical extension of the feasibility experiments, and if successful, a transition of this research would be tested in clinical trials

Award # 5R01DK067683-04 Agency NIH Lead Institution University of Texas-MD AndersonTypeExtramural

Explanatory Notes The goal of this project is to use nanoshells for enhanced MR imaging. The EHS-relevant component of this research is the mechanistic understanding of cell uptake of nanoparticles.

Abstract We hypothesize that specific vascular addresses within tumor vasculature can be exploited for imaging and detection of metastatic breast carcinoma; our goal is to use these biochemical differences to develop targeted therapies. Here, we propose to investigate the molecular diversity of angiogenic vasculature during the tumor progression and metastases of breast cancer. Our specific aims are (i) to identify and characterize suitable markers of bone marrow metastases as targets for vascular imaging; (ii) to study the localization and distribution of the probes and respective receptors by imaging systems; (iii) to design, synthesize and validate devices for targeted imaging by developing novel tools for intravital imaging at the protein-protein level (such as engineered phage particles, recombinant proteins, nanoshells, or fluorescent microspheres). The most efficient targeting systems will be tested and validated in vivo in mouse models of bone marrow metastases. If successful, novel strategies to image metastatic breast cancer will be derived from this application. The approaches utilized in this application can be used to characterize the tumor microenvironment in breast cancer, changes and localization of receptors in the vascular endothelium of tumor blood vessels during breast cancer progression. In addition, probes that target breast cancer vasculature will be developed as delivery tools and will likely enhance effectiveness of current imaging technology

Award # 1Z01DK043400-07 Agency NIH Lead Institution TypeIntramural

Explanatory Notes The EHS component of this project is the mechanistic understanding of nanoparticle uptake, distribution, and clearance by using gadolinium nanoparticles for imaging. The EHS-relevant component of this research is

Abstract Renal disease is difficult to detect, particularly in a form called acute kidney injury (also known as acute renal failure). We are developing new methods to detect renal disease involving either MRI, or urine or blood tests. 1) Detection of proximal tubule damage in mice MRI using dendrimer gadolinium chelate nanoparticles. We found that Gadolinium nanoparticles accumulate in the proximal tubule, and can be used to detect renal structure, function, and injury. We extending our methods for the early detection and outcome prediction of sepsis-acute kidney injury to other forms of kidney damage, including chronic kidney disease. 2) Markers for early diagnosis. We are new using microarray and proteomic techniques to search for early biomarkers of ischemia/reperfusion-, nephrotoxic-, and sepsis-induced acute kidney injury. We have a few excellent candidates, including Fetuin A, that are being validated using our mouse and rat acute kidney injury models. 3) We have worked out the collection, storage, and processing conditions to use urinary exosomal proteins as biomarkers of renal disease. We are beginning to search for exosomal markers of structural renal injury in our animal models

Award # 5R01NS034608-10 Agency NIH Lead Institution Oregon Health And Science UniversityTypeExtramural

Explanatory Notes The goal of this project is to use iron oxide nanoparticles for enhanced MR imaging. The EHS-relevant component of this research is the mechanistic understanding of influx and uptake of nanoparticles to the brain.

Abstract The efficacy of gene therapy against brain tumors will depend upon delivery of viral vector throughout tumor and specific cytotoxicity toward both infected and non-infected tumor cells. Previously this project has focused on delivery, assessing interstitial infusion and transvascular delivery of particles by osmotic opening of the blood-brain barrier (BBB). While delivery remains our major focus, the current proposal will also move to a broader examination of virus and particle uptake and efflux. In addition to recombinant adenovirus vectors, as a model for virus particles we will use viral-sized iron oxide nanoparticles, Combidex and Code7228, because they allow direct comparison of magnetic resonance (MR) imaging with histology and ultrastructure. In Aim 1 we will assess influx and uptake of virus and iron particles into rat brain and intracerebral tumor, and investigate the effect of tumor size, permeability and prior irradiation. Since radiation may increase virus distribution and/or transgene expression, we will test a novel approach combining tumor-specific radioimmunotherapy with virus delivery. This aim will also evaluate iron particle influx in a rat stroke model, and characterize phagocytic and/or astrocytic reactive cells responsible for iron particle trapping. Aim 2 will evaluate virus and iron particle efflux from the brain, using mR and histology to delineate efflux pathways. In Aim 3, we will investigate the potential for gene therapy with Herstatin, a secreted protein which inhibits the epidermal growth factor receptor (EGFR). We will compare intratumor vs. transvascular delivery of protein and a Hestatin adenovirus construct, both in the LX-1 lung cancer metastasis model, as well as in glioma and breast metastasis models. We hypothesize that because Herstatin is secreted and has high activity against EGFR overexpressing cells, it will provide bystander efficacy even when a small proportion of tumor is infected. Finally, Aim 4 will be an expanded clinical protocol of rion particle imaging and localization, to assess BBB and blood-tumor barrier permeability and Combidex uptake in adult and pediatric brain tumors, and in CNS inflammatory lesions. We will also evaluate tumor vascularity using MR angiography with Code7228, a new formulation which allows bolus administration and dynamic MRA superior to Gd. Our hypothesis is that tumor imaging with the iron oxide particle agents requires both a leaky BBB and trapping by uptake into reactive cells. We anticipate that these studies will not only be useful in designing clinical trials of brain tumor gene therapy, but also in providing a new means to image CNS tumors, neurological lesions and even gene therapy approaches which evoke a cellular reaction

Award # 5R21NS052030-02 Agency NIH Lead Institution University of WashingtonTypeExtramural

Explanatory Notes The EHS component will characterise the interaction of nanoparticles with neurons.

Abstract The ability to introduce exogenous nucleic acids to cells in the central nervous system (CMS) is a powerful technique with applications in neurobiology research and in treatment of neurological disease. RNA interference (RNAi) is currently the most potent and specific method for blocking gene expression; however, there is not to date a successful in vivo application of RNAi to a mammalian model of neurological disease. The major challenge to realizing the full therapeutic value of RNAi for the CMS lies in the delivery of the nucleic acids to target cells. Synthetic nanoparticles offer in vivo protection, low immunogenicity, and relative ease of manufacturing and scale up, and have been used to deliver plasmid and oligonucleotides to various types of cultured cells. However, successful application of this technology for neuronal cell delivery both in vitro and in vivo has been limited due to low delivery efficiencies. The major goal of this research proposal is to develop nanoparticles that mediate efficient neuronal delivery of short, interfering RNA (siRNA), molecules that mediate RNA interference. We propose to implement a novel strategy to overcome intracellular transport barriers by designing nanoparticles that "hitchhike" on motor proteins that transport vesicles toward the cell body. This goal can be achieved by realizing the following aims: (i) synthesizing nanoparticle formulations that integrate components for neuron targeting, vesicle release, and motor protein-assisted, retrograde transport, and optimizing formulations by evaluating the delivery efficiency in postmitotic neuron-like PC 12 cells, (ii) demonstrating siRNA delivery and specific downregulation of transgene expression in primary neurons, and (iii) achieving nanoparticle delivery to neuronal cell bodies in the brain by retrograde transport from spinal cord injection. Efficient delivery systems for the CNS are crucial for both research and clinical applications; thus, successful completion of this project would result in a major step toward realizing the full potential of this technology.

Award # 5R01NS050660-02 Agency NIH Lead Institution University of Nebraska Medical CenterTypeExtramural

Explanatory Notes The goal of this project is to use Nanogel for targeted drug delivery across the blood/brain barrier. The EHS-relevant component of this research is the mechanistic understanding of brain uptake of nanoparticles.

Abstract Permeability of various drugs across the blood-brain barrier (BBB) is significantly dependent on the expression and functional activity of specific efflux transporters located in the membrane of brain capillary endothelial cells (BCEC). Selective transient downregulation of the transporters will lead to the application of more effective and less toxic doses of therapeutic drugs against brain tumors or viral infections in CNS. Previously, antisense inhibitors have been shown to temporarily arrest the synthesis of major multidrug resistance agent, membrane P-glycoprotein, and promote reversal of the resistant cell phenotype. The more effective RNA interference mechanism has been recently discovered for selective switching off expression of various genes. Short hairpin RNA (siRNA) could be introduced into target cells through a plasmid DNA precursor using methods of non-viral gene therapy. However, targeted delivery of the pDNA to the cells of the BBB requires a good systemic carrier and selective vectors that bind to the BCEC. As such a carrier, polymer crosslinked Nanogel particles modified with the brain-specific homing peptides (BSHP) have been chosen for tranfection of the BBB by shRNA-encoding plasmid DNA with an ultimate goal suppressing the specific membrane proteins, drug efflux transporters, in the BBB. Specific BSHPs to be attached to the surface of the Nanogel and target delivery to the BBB have been selected in vivo from a vast amount of peptides in the phage display library. Nanogel is non-toxic and highly effective as a transfection agent in many cell lines and evidently, one of the carriers with great potential for systemic administration. The vectorized RNA Interference-Producing system (RIP system) could be used for bioengineering of the BBB permeability for therapeutic agents whose brain accessibility was hampered by specific drug efflux transporters. The central hypothesis of the proposal is that transient suppression of selected drug efflux transporters in the BBB via systemic transfection of brain endothelium using targeted RNAi-producing systems can result in significant enhancement of drug transport to the brain during chemotherapy of the CNS-related diseases. Our Specific aim 1 is to develop the BCEC-targeted Nanogel carriers for systemic delivery of plasmid DNA to the BBB. Specific aim 2 is the enhance transfection efficacy of the BCEC-targeted Nanogel carriers in vitro and in vivo. Specific aim 3 is to suppress selected drug efflux transporters in the BBB in vivo and temporary increase drug transport into the brain. In this Aim brain transport of several representive nucleoside analogue drugs will be assessed in animal model following the transient downregulation of drug efflux transporters in the BBB by the Nanogel-based RNAi-producing systems

Award # 5P42ES013660-030002 Agency NIH Lead Institution Brown UniversityTypeExtramural

Explanatory Notes Studies of mixed dust including inert and metal-bearing nanoparticles will aid in the development of rapid screens for toxic dusts.

Abstract Dusts generated during demolition of buildings, remediation, and construction may be contaminated with toxic particulates including metals and asbestos fibers. Three forms of asbestos are on the CERCLA Priority List of Hazardous Substances and it is a common contaminant at former industrial sites and military bases. However, the potential adverse health effects resulting from inhalation of mixed dusts from these asbestos-contaminated sites are unknown. The availability of transition metals, especially iron, to participate in redox cycling leading to generation of free radicals has been hypothesized to play an important role in asbestos-induced lung diseases. It is hypothesized that contamination with iron and asbestos fibers will potentiate the toxicity of mixed dust exposures due to enhanced iron mobilization, redox cycling, and generation of reactive oxygen species. The ultimate goal of this project is to develop a panel of cellular and molecular endpoints that can be integrated into a short-term screening strategy to identify toxic mixed dusts. The Specific Aims to achieve this goal are: a) To generate and characterize model dust samples composed of inert particles, metal-contaminated particles, and asbestos fibers; b) To assess mobilization of transition metals and redox activity of these model dust samples in vitro; c) To develop a gene expression profile that will differentiate between nontoxic and toxic dusts; d) To explore the mechanisms of mixed dust-fiber toxicity using short-term in vitro assays; and e) To validate this testing strategy in a chronic carcinogenicity assay

Award # 0646507 Agency NSF Lead Institution University of Minnesota-Twin CitiesTypeExtramural

Explanatory Notes

Abstract It is hypothesized that lung deposition of highly agglomerated nanoparticles differs from that for spherical particles of the same mobility size (diffusivity). We will synthesize nanoparticulate agglomerates that are similar in composition and structure to those that are currently being produced in large volumes, and we will accurately measure their transport properties (diffusion coefficient, sedimentation speed, dynamic mobility,
aerodynamic diameter) as a function of their mobility-equivalent diameter. We will then
measure the mobility-size-dependent deposition of these particles in models of the respiratory system extending from the mouth through the 8th generation of the upper respiratory tract. Deposition of spherical nanoparticles as small as 3 nm in the same airway models will be measured, and these results will be compared with values for the agglomerates to determine whether or not agglomerate structure affects deposition in this part of the respiratory system.

Award # 0553682 Agency NSF Lead Institution University of Minnesota-Twin CitiesTypeExtramural

Explanatory Notes This NSF project contains funding aimed at understanding and designing nanostructured interfaces for drug delivery targeting that specifically binds to infection or inflammation sites in human body. This engineered formulations of fractalkine-targeted stealth liposomes, their transport properties and binding capabilities will be evaluated in vitro.

Abstract Currently, the main problems associated with systemic drug administration are the necessity of a large drug dose to achieve high local concentration, non-specific toxicity, and other adverse side-effects due to high drug doses. Targeted drug delivery can bring a solution to all these problems. With the emergence of stealth liposomes (liposomes covered with polyethylene glycol), the use of liposomes as drug delivery vehicles has received a new impetus, and several successes have been reported. In order to further improve upon different therapies, clinically active stealth liposomes need to include site-directed ligands to enhance their specificity for the pathological site. Peptides that recognize specific cell types or specific macromolecules on the cell surface can serve as targeting agents.

The project focuses on engineering peptide-amphiphiles, and designing nanostructured interfaces that specifically target fractalkine, a novel adhesion molecule on the surface of endothelial cells that is expressed only at sites of infection or inflammation. The hypothesis of this research is that fractalkine can serve as a specific target moiety for drug delivery targeting. The PIs group is the first one that is engineering fractalkine-targeted drug delivery systems. The proposed approach will utilize fractalkine as of the N-terminus of the fractalkine receptor (NTFR peptide-amphiphile), as the bullet. Preliminary work in the PIs lab has demonstrated that liposomes functionalized with the NTFR bind preferentially to inflamed human umbilical vein endothelial cells (HUVECs) in a concentration dependant manner. In addition, targeting of liposomes to inflamed HUVECs over healthy HUVECs is significantly increased when two adhesion receptors are employed, and the interface of the liposome is functionalized with NTFR that binds to fractalkine, and a second peptide-amphiphile that binds to 5 1 integrin. This one year project will engineer different formulations of fractalkine-targeted stealth liposomes, with nanostructured interfaces composed of peptide-amphiphiles that bind to the target(s), and polyethylene glycol of varying density and molecular weight. The designs will be evaluated in vitro in terms of binding, specificity, and internalization efficiency.

Broader Impacts of the Proposed Research
Fractalkine has been detected in a variety of diseases such as: cardiac allograft rejection; prostate, lung, and colorectal cancer; pulmonary arterial hypertension; AIDS; atherosclerotic coronary artery disease, the leading cause of death in the USA; rheumatoid arthritis; and other inflammatory conditions. Therefore, the selection of fractalkine as a target for drug delivery is of great therapeutic value. This project will be the seed for future interdisciplinary research at the interface of synthetic chemistry, biology, nanotechnology, and engineering, with applications such as targeted drug delivery, and biosensors.

Award # 0602906 Agency NSF Lead Institution North Carolina State UniversityTypeExtramural

Explanatory Notes

Abstract We have assembled a multidisciplinary team of Russian and U.S. researchers who are experts in experimental/computational Materials Science and Medical/Agricultural Research. This team will create hydrosols of functionalized diamond nanoparticles that act as designer gastrointestinal enterosorbents for mycotoxins. Mycotoxins, which are the toxic by-products of molds, are a major problem in developing countries where food shortages make disposing of potentially moldy feed impractical. The designer diamond nanoparticles being developed by our team are a completely new and novel set of enterosorbent materials that can be made in bulk quantities and tuned to bind specific toxins while minimizing harmful side effects. The U.S. team is composed of Prof. Don Brenner and Prof. Tzy-Jiun Mark Luo from the Department of Materials Science and Engineering at North Carolina State University, and Dr. Olga Shenderova, Senior Scientist and Dr. Varvara Grichko, Consultant, from the International Technology Center (ITC), Research Triangle Park, NC. The primary role of the U.S. researchers is to use accurate ab initio and molecular modeling methods to efficiently explore diamond nanoparticle surface structures in terms of stability and target multi-functionality, and using a proprietary glow discharge atmospheric plasma experimentally process and then spectroscopically characterize diamond nanoparticles based on the results of the modeling studies. Multi-functionality refers to the ability of the diamond nanoparticles to remain as stable and well dispersed hydro/organosols while yielding biological activity for targeted anti-toxin applications. The lead researchers of the Russian counterpart effort are Dr. Vladimir Bondar and Dr. Alexei Puzur of the Institute of Biophysics (IB), Russian Academy of Sciences, Siberian Branch, Krasnoyarsk, Russia. Dr. Bondars and Dr. Puzurs primary role in our team is to provide size-selected diamond nanoparticles for processing at the ITC, and to carry out the in vitro and in vivo studies of the biological activity, both desirable and harmful, of the materials after processing by the U.S. team. This institute is one of the international leaders in the development and testing of enterosorbents for livestock and human medical uses. As part of this effort we are training a new generation of Materials Science graduate students to perform research at the interface of Materials Science/Medicine/Agriculture, as well as providing these students first-hand experience with how theory, modeling and experiment can interact and aid progress in each area. This effort also gives students experience working within an industrial setting and provides the students with intimate knowledge of technology transfer.

Award # 5R01CA098194-04 Agency NIH Lead Institution North Carolina State UniversityTypeExtramural

Explanatory Notes

Abstract This proposal describes the synthesis of a general class of nanoparticle delivery vectors based upon hybrid biomolecule-gold nanoparticle complexes. The vectors are designed to transport therapeutic oligonucleotides across cell membranes to target cancer cell nuclei. Therapeutic oligonucleotides are synthetic single stranded nucleic acid molecules that are resistant to digestion by nucleases. A multifunctional approach has been developed which combines cell-specific recognition ligands, endosomal escape peptides, nuclear localization signals and controlled release of therapeutic oligonucleotides to cells. The ability to target biomolecule-nanoparticle complexes to specific cell types, through a relatively simple attachment of cell-specific ligands or peptides, provides the potential for diagnosis and treatment of cancer. A quantitative test for nuclear delivery of an oligonucleotide drug to the nuclei of HeLa cells is proposed as an assay for the efficiency of delivery of a therapeutic agent. The long term goal of this research is to develop a non-viral vector capable of delivering therapeutic oligonucleotides to cancer cells in vivo. Achieving this long term goal requires the following steps. 1. Formulate stable nanoparticle-bioconjugates that are resistant to aggregation and chemical exchange in biological fluids and cells. 2. Develop microscopy techniques to monitor nanoparticle trajectories and quantitate localization in cells. 3. identify the best combination of peptides for performing the functions of endocytosis, endosomal escape and nuclear uptake. 4. Characterize fundamental interactions of protein-peptide conjugates with nanoparticles including their susceptibility to exchange or replacement. 5. Develop strategies for covalent attachment of proteins and oligonucleotides to nanoparticles. 6. Determine of the efficiency of regulated in vitro protein expression using a well-characterized model system

Award # 5R01GM063679-04 Agency NIH Lead Institution University of FloridaTypeExtramural

Explanatory Notes

Abstract Local anesthetics (LAs) reversibly prevent impulse transmission in nerves by voltage-, time- and frequency-dependent blockade of sodium channel conductance (I-Na). These pharmacological actions underlie the therapeutic use of LAs in clinical care to provide regional anesthesia (e.g., epidural blockade) for patients undergoing surgical procedures or childbirth. However, inadvertent intravascular injection or overdose may lead to undesired INa blockade in other tissues (e.g., heart, central nervous system) and thereby cause potentially life threatening adverse events (e.g., cardiac arrest, seizures). Although seizure activity and respiratory depression caused by LA overdose are potentially life threatening, these events can be readily treated with antiepileptic medications and controlled artificial ventilation, respectively. Of equal or greater importance, few options (e.g., ACLS) currently exist for treatment of cardiac toxicity caused by intravascular injection. For these reasons, an agent or technique allowing rapid, efficacious treatment of the cardiac effects of LA toxicity would be useful. The objectives of this grant are to generate the knowledge necessary to create agents specifically designed to treat patients suffering from the toxic effects of LAs. Recent advances in particle science engineering now afford new and exciting opportunities to develop highly effective therapeutic strategies aimed at successfully treating drug poisonings. Specifically, the recent advent of nanotechnology with its tremendous potential to solve major biomedical problems now offers unparalleled opportunities to solve the problem of LA toxicity. Four types of biocompatible and biodegradable nanoparticles (NPs) with 10-100 nm diameter will be synthesized by colleagues in the NSF Engineering Research Center for Particle Science and Technology for detoxification of LAs. These NPs will rely on absorption (microemulsions), adsorption (electron acceptor), or both mechanisms (2 types of "smart" microemulsions) to reduce the free concentration of LA in various media and decrease the biological effects of LA in tissues and intact organisms. The NPs will be studied to 1) detail the physicochemical characterization of the NP-LA interaction (Objective A), and 2) determine whether the cardiotoxic effects of LAs can be attenuated by NPs in biological systems (Objective B). This highly multidisciplinary project spanning organic chemistry, engineering, and medicine contains two objectives and three specific aims: Specific Aim #1: Determine the extraction efficiency of NPs to remove LAs from simple (normal saline) and complex (human plasma and blood) media. Optimize LA extraction efficiency of the various NPs. Specific Aim #2: Determine the molecular mechanisms whereby the different types of NPs can efficiently extract LAs. Specific Aim #3: Determine the effectiveness of nanoparticles to attenuate or reverse the cardiotoxic effects of LAs at three functional levels: 1) single cell (ventricular myocytes), 2) tissue (isolated hearts), and 3) intact rat (closed chest).

Award # 5R44GM063283-04 Agency NIH Lead Institution SUNY at Stony BrookTypeExtramural

Explanatory Notes This EHS component of this grant focuses on developing anti-adhesion nanostructured products that reduce the formation of internal adhesions after surgery by assaying for biocompatibility of these nanostructured products using in vitro methods.

Abstract Stonybrook Technology and Applied Research (STAR), Inc. is currently focused on the development of unique anti-adhesion nanostructured products in post-operative surgery. Adhesions are mainly induced from the trauma of surgery and can lead to serious complications, including pelvic pain, small bowel obstruction, female infertility, chronic debilitating pain and difficulty with future operations. Typically, a patient will often undergo surgery to remove adhesions, only to have them reform. It is a very costly problem, e.g., Medicare alone has paid $3.22 billion per year for the treatment of adhesion related complications in the 1990's. In this SBIR Phase II submission to the NIGMS, STAR is seeking 3 years of support to complete the development of two FDA-regulated anti-adhesion products: (1) unsupported membranes for abdominal surgery and (2) mesh supported membranes for hernia repair. The specific aims of this application are: 1. Thorough in vivo studies using rat and rabbit models to evaluate the proposed two anti-adhesion products. 2. Introduction of medicated alternatives and composition adjustments as contingency plans. 3. Development of mass production facility using multiple-jet electro-spinning technology to commercialize anti-adhesion products. Thus, in the Phase II project, the proposed tasks will deal mainly with the completion of: 1. In vivo studies using the objective rat model and the rabbit model to evaluate the anti-adhesion properties of two electro-spun non-medicated products based on a successful chemical composition. 2. Optimization of non-medicated anti-adhesion products by fine tuning of the PEG composition using in vitro and in vivo evaluations as a first contingency plan. 3. Optimization of medicated anti-adhesion products by determining the minimum effective dosage of Cefoxitin sodium for local delivery based on in vivo evaluation as a second contingency plan. 4. Development of scale-up instrumentation by the implementation of electro-blowing technology, robust linear fluid delivery system, uniform thickness production and system integration

Award # 2R44GM072142-02 Agency NIH Lead Institution Nanomedex, IncTypeExtramural

Explanatory Notes The goal of this grant is to make a nanoparticle-based emulsion of the propofol, a widely used anesthetic. The EHS-relevant component of this research is to determine the pharmacokinetic effects and biocompatibility properties of this emulsion using in vivo model systems.

Abstract Propofol is the largest selling, intravenous general anesthetic with domestic annual sales in excess of $500 million due to several favorable characteristics of this anesthetic and has an expected annual growth rate of 19%. However, a primary drawback of propofol (2,6-diisopropylphenol) revolves around this drug's extreme lipophilicity that necessitates dispersion in a soybean macroemulsion (i.e., Intralipid 10%). This requirement causes several possible adverse drug outcomes (e.g., rapid bacterial growth, severe stinging pain on injection). In SBIR Phase I activities, NanoMedex proposed that these adverse reactions caused by the Intralipid formulation could be prevented by using an alternative formulation of microemulsion-based nanoparticles to construct clear, thermodynamically stable formulations of propofol. To that end, NanoMedex demonstrated in SBIR Phase I the physical boundary parameters to synthesize these formulations, that these formulations had differential release rates in response to dilution, that these NanoMedex propofol formulations had differential anesthetic kinetics in rat, and that one NanoMedex formulation and Diprivan (the commercial formulation) were bioequivalent to cause anesthesia in dog. In SBIR Phase II, NanoMedex seeks to expand on these findings to demonstrate the following Specific Aims leading to an US Food and Drug Administration Investigational New Drug application and GMP-grade propofol formulations. Specific Aim 1. Determine the propensity of propofol microemulsions (NMDX) and a macroemulsion (Diprivan) to cause stinging on injection into a rat tail vein. Specific Aim 2. Characterize the ability of microbes to survive and grow in different anesthetic solutions (NMDX formulations, Diprivan, Intralipid, and Baxter PPI). Specific Aim 3. In swine, determine the pharmacokinetics of a propofol bolus and infusion as well as and dose-response relationship to cause hemolytic and/or thrombotic changes due to propofol microemulsions (NMDX) and macroemulsion (Diprivan). Specific Aim 4. Determine the stability of a GMP-manufactured, NanoMedex propofol microemulsion and macroemulsion (Diprivan) to changes in time, light, temperature, oxygen, and vibration. GMP-grade propofol formulations will be custom-synthesized in an FDA-approved plant with appropriate regulatory oversight using external contractors. At the conclusion of SBIR Phase II, NanoMedex, Inc. will have applied for an FDA IND, secured external financing via private investors to conduct FDA Phase I studies, and begun planning for an abbreviated new drug application. Successful conclusion of the propofol project will allow NanoMedex, Inc. to present this propofol microemulsion to large Pharma for additional Phase ll/lll studies and commercialization.

Award # Agency NIH Lead Institution TypeExtramural

Explanatory Notes

Abstract National Toxicology Program

Award # Agency NIOSH Lead Institution NIOSH/HELDTypeIntramural

Explanatory Notes

Abstract This project will employ in vitro and in vivo models to determine if particle surface area is a more predictive metric of toxic exposure than particle mass. The toxicity of ultrafine particles from welding fumes, diesel exhaust and combustion, and special purpose nanoparticles is of concern. This project addresses two major issues. (1) Is particle surface area a more appropriate dose metric than mass when attempting to understand the toxicity of ultrafines? (2) Can in vitro models become more predictive of in vivo response with the use of a particle surface area per exposed cell surface area metric of exposure? These data will be used to model lung burden vs. response. Such identification of the proper metric for exposure would be invaluable to risk assessment for ultrafine particles.

Award # 1R01OH009141-01 Agency NIOSH Lead Institution Ohio State UniversityTypeExtramural

Explanatory Notes

Abstract The objectives of this program are to verify two hypotheses. First, the quantifiable differences in surface reactivity of nanoparticles, as measured by acidity, redox chemistry, metal ion binding and Fenton chemistry as compared to micron-sized particles of similar composition cannot be explained by the increase in surface area alone. Second, the oxidative stress and inflammatory response induced by nanoparticles upon interaction with macrophages and epithelial cells is dependent on their surface reactivity. The basis of these hypotheses is that nanoparticles contain significantly higher number of "broken" bonds on the surface that provide different reactivity as compared to larger particles. The experimental approach focuses on three classes of manufactured nanoparticles, catalysts (aluminosilicates), titania and carbon. For the catalysts and titania samples, nanoparticles (< 100 nm) and micron-sized particles of similar bulk composition will be studied. For carbon, carbon black and single walled carbon nanotubes are chosen. Nanoparticles of aluminosilicates and titania will be synthesized, whereas the other particles will be obtained from commercial sources. Characterization will involve electron microscopy, surface area, surface and bulk composition. Reactivity of well- characterized particles in regards to their acidity, reaction with antioxidants simulating the lung lining fluid, coordination of iron and Fenton chemistry will be carried out using spectroscopic methods. Particular attention will be paid to surface activation as may exist during manufacturing and processing. In-vitro oxidative stress and inflammatory responses upon phagocytosis of the particles by macrophages and pulmonary epithelial cells will form the toxicological/biological end points of the study. Methods include gene array techniques, assays for reactive oxygen species and adhesion molecules on endothelial cells. As nanotechnology advances are made over the next decades, exposure to nanoparticles is going to increase significantly. Potential inhalation risks exist during manufacture, product recovery, processing and in some cases during consumer use. Risk assessment and management will be facilitated if broad generalizations can be developed correlating surface structure and reactivity of nanoparticles, much like we classify molecules and their toxicity. The expected results from this project will examine for the first time how unsaturated bonds on nanoparticles influence their chemical and biological reactivity. Preparation of nanoparticles in an activated state with unpassivated surface atoms is an important part of this research program

Award # Agency NIOSH Lead Institution NIOSH/EIDTypeIntramural

Explanatory Notes

Abstract Eileen Kuempel is collaborating with the Chemical Industry Institute of Toxicology Centers for Health Research on software modifications for use in lung dosimetry modeling.

Award # Agency NIOSH Lead Institution NIOSH/DARTTypeIntramural

Explanatory Notes

Abstract There is mounting evidence that the toxicity of some aerosols may be closely associated with the number or surface-area of inhaled particles. Low-solubility ultrafine (typically smaller than 100 nm) and high specific area particles are of particular concern. This project is part of a wider research program aimed at studying te toxicity of work-place related aerosols within this category, including those associated with nanotechnology. Methods will be developed to generate and deliver well-characterized particles to exposure systems, enabling particle characteristcs responsible for specific toxic responses to be investigated in a systematic manner. The research will include the development of off-line and on-line aerosol and particle characterization techiniques, including methods to measure aerosol surface-area, and methods to characterize the composition and structure of nanometer-diameter particles.

Award # 0507036 Agency NSF Lead Institution Old Dominion University Research FoundationTypeExtramural

Explanatory Notes

Abstract The proposed work involves the design of nanoparticle probes to study the dynamics of "smart" membrane transport and the size change of membrane pores of living bacterial cells in real time. The model system to be used is the efflux pump (MexA-MexB-OprM) in P. aeruginosa because it extrudes a wide range of structurally and functionally unrelated substrates. The primary research goals are to (1) study the dependence of membrane transport kinetics on the size (1-80 nm), shape (sphere, rod, or triangle), surface charge (negative vs. positive), and properties (hydrophobic vs. hydrophilic) of nanoparticles, (2) study the interactions and assembly of efflux pump proteins in real time in response to the presence of synthesized nanoparticles with different sizes, shapes, surface charges, and properties, and (3) study the cytotoxicity and genotoxicity of the synthesized nanoparticles in vitro and in vivo to identify biocompatible and environmentally-friendly nanoparticles for real-time probing of membrane transport in living cells. This is an interdisciplinary research project in the areas of chemistry, material science and engineering, molecular and cellular biology, and protein engineering involving 4 PIs from Old Dominion University and Northwestern University: Xu (ODU, Chemistry and Biochemistry), Elsayed-Ali (ODU, Electrical and Computer Engineering), Osgood (ODU, Biological Sciences), and Van Duyne (Northwestern, Chemistry and Materials Science). An additional PI from CEA in France (Gillet, Protein Engineering) is also involved.

Award # 0609311 Agency NSF Lead Institution University of FloridaTypeExtramural

Explanatory Notes

Abstract This project will determine if adhesive interactions can be used to predict and isolate potentially beneficial or deleterious nanostructures for specific biological activity. It would be the first attempt to directly measure interaction forces between living cells and engineered nanostructures. This new cellular probe force microscopy (CPFM) technique will attach tissue culture to a probe that can assess large regions in a single experiment. Preliminary results show that the CPFM technique is sensitive to directly probe cell receptor-ligand binding. The project work plan was well presented with good justifications for each of the main experimental components. These justifications for experimental simplicity do eliminate more environmentally relevant research, however, the aim of the project is to demonstrate the feasibility of the tool which can then be applied to many different fields.

Award # 0630823 Agency NSF Lead Institution Clemson UniversityTypeExtramural

Explanatory Notes

Abstract Nanomaterials and their broad implications are rapidly reshaping the landscape of our science and technology. This proposed exploratory research is to address the crucial need of understanding lifeless nanomaterials in biological systems and in the environment. Three objectives are designed to examine the biophysical interactions of single-walled carbon nanotubes (SWNTs), a major class of nanomaterials, and naturally occurring lysophospholipids, and to investigate the toxicity of lipid-solubilized SWNTs in cells and in whole animals. These objectives will be investigated by a synergetic team of researchers who have track records of creativity and productivity and whose expertise encompasses the fields of nanotechnology, biophysics, biology, engineering, and environmental toxicology.

Award # 0425626 Agency NSF Lead Institution Ohio State UniversityTypeExtramural

Explanatory Notes The goal of this center is to design and fabricate nanofluidic circuits for manipulating individual biomolecules. The EHS focus of this work lies in designing and assessing transport and biocompatibility of various nanostructures in the test system.Biocompatibility issues will be addressed in parallel with the development of new nano-fluidic designs and devices.

Abstract The Nanoscale Science and Engineering Center entitled Center for Affordable Nanoengineering of Polymer Biomedical Devices (CANBD) is a partnership between the U. of Akron, Boston University, UC Berkeley, Johns Hopkins, Florida A&M, and Purdue. The NSEC includes 38 investigators from 9 departments.

The Center seeks to develop polymer-based low-cost nanoengineering technology that can be used to produce nanofluidic devices and multifunctional polymer-nanoparticle-biomolecule nanostructures for the next generation medical diagnostic and therapeutic applications. The research plan is comprised of three thrust areas. The Nanomanufacturing Thrust Area combines 'top-down' fabrication and 'bottom-up' molecular self-assembly to produce well-defined passive and active nanostructures. In the Transport Phenomena Thrust Area, the research will achieve design capabilities at the nanoscale by combining nanofluidic design, transport phenomena at the nanoscale, and multiphase transport structures with multiscale modeling and macroscalar property assessment. Biocompatibility issues will be addressed in the Biocompatibility Thrust Area in parallel with the development of new nanofluidic designs and devices.

The near-term goal of the three closely linked research thrust areas is to design and fabricate polymer-based, 3D nanofluidic circuits for manipulating the shape, orientation and transport behavior of individual biomolecules in well-defined nanoscale flow fields (5-100 nm). Test bed examples include a simple, handheld protein separation/diagnostic device; a nanoneedle cell patch for low-invasive delivery of genes and macromolecular medicines into cell walls; and biomolecular nanopumps as synthetic ion channels. The ultimate goal is to design and assemble a nanofactory based on the integration of nanofluidic circuits, synthetic chemistry and biological complexation.

Center collaborators include at least 20 companies in Ohio and the U.S., Battelle, the Cleveland Clinic Foundation, the National Cancer Institute, Oak Ridge National Laboratory, Wright Patterson Air Force Labs, and researchers in Asia, Australia and Europe. The Center also plans to coordinate closely with NSECs at the University of California at Los Angeles and the University of Illinois-Urbana (nanomanufacturing), the NSF STC at the University of North Carolina at Chapel Hill (environmentally responsible solvents), and the NSF ERCs at the University of Washington (biomaterials and biocompatibility) and the Georgia Institute of Technology (3D tissue models) because of complementary research agendas.

The education and outreach vision of the Center is to integrate the latest research developments into a practical student curriculum that imparts multidisciplinary skills and global awareness to both graduate and undergraduate students. The key education elements include a series of new courses to introduce nanoengineering of biomedical devices and related topics; an interdisciplinary curriculum offering an undergraduate minor and a graduate certificate; internships and visits to industry and national laboratories in the U.S. and abroad; and web-based dissemination. The recruitment and retention of minorities and women will be emphasized through close collaboration with minority institutes such as FAMU/FSU. Undergraduate students will participate in research via senior honors theses and targeted REU support. Outreach activities include web-based science modules for K-12 students nationwide; workshops and short courses for high school science teachers and industrial researchers; and on-site research projects and workshops for middle school and high school students supervised by graduate students

Award # RD-833320 Agency EPA Lead Institution Oregon State UniversityTypeExtramural

Explanatory Notes

Abstract Rapid growth of the nanotechnology industry is resulting in increased exposure of humans and the environment to nanomaterials prior to the scientific investigation of potential risks. It is clear that there is a need to develop rapid, relevant and efficient testing strategies to assess these emerging materials of concern. Here we propose an in vivo system for rapidly assessing the toxicity of nanomaterials at multiple levels of biological organization (i.e. molecular, cellular, systems, organismal). Early developmental life stages are often uniquely sensitive to environmental insult, due in part to the enormous changes in cellular differentiation, proliferation and migration required to form the required cell types, tissues and organs. Molecular signaling underlies all of these processes. Most toxic responses result from disruption of proper molecular signaling, thus, early developmental life stages are perhaps the ideal life stage to determine if chemicals or nanomaterials are toxic. Our hypothesis is that the inherent properties of some engineered nanomaterials make them potentially toxic. To test this hypothesis we specifically propose to: (1) Further develop our in vivo zebrafish toxicity assay to define the in vivo responses to nanomaterials (2) Begin to define structural properties of nanomaterials that lead to adverse biological consequences.
Approach: We propose a three-tier approach exploiting the advantages of the embryonic zebrafish model to assess the toxicity of nanomaterials. Tier 1: Rapid screening experiments will be conducted to assess the toxicity of a wide range of structurally well-characterized nanomaterials commercially available or produced by researchers of the Oregon Nanoscience and Microtechnologies Institute (ONAMI). Nanomaterials found to elicit significant adverse effects will proceed to Tier 2 testing. Tier 2: Potential cellular targets and modes of action will be defined in vivo using a suite of transgenic fluorescent zebrafish and indicators of cellular oxidative state. Nanomaterials will be grouped according to structural indices and effects. Representative nanomaterials from each group will be selected for Tier 3 testing. Tier 3: Global gene expression profiles will be used to define the genomic responses to nanomaterials. Data from these studies will be used to define structure-activity relationships using a Nanomaterials Effects Database we have created to collate, organize and analyze data on nanomaterial effects across species and exposure scenarios.
Expected Results: The successful completion of these studies will fill important gaps in our understanding of the human health risk posed by exposure to nanomaterials. The proposed research will deliver: (1) a validated in vivo system for rapidly assessing existing and future novel nanomaterials (2) data on nanomaterial structure effects relationships.
Supplemental Keywords: dose-response, teratogen, animal, stressor, toxics, particulates, nanotechnology, nanotoxicology, environmental chemistry, Northwest, Oregon, OR, industry

Award # 5R01CA102791-04 Agency NIH Lead Institution University of Nebraska Medical CenterTypeExtramural

Explanatory Notes The goal of this project is to develop targeted nano-probes for molecular imaging to enable non-invasive early detection of cancer.  The EHS relevant component of this research involves the pharmacokinetic testing of nano-probes (linking peptide targets to a variety of nanostructures) in mouse models to identify the best candidate nanoprobes for clinical evaluation.

Abstract Effective in chemotherapy of cancer and viral infections nucleoside analogues (NA) are actually 'prodrugs', which must be first converted in vivo into nucleoside 5'-monophosphates and, finally, into the drug's active form, nucleoside 5'-triphosphates. They efficiently terminate DNA synthesis and are cytotoxic for the proliferating cancer cells. However, therapeutic NAs in the form of 5'-triphosphates are considered too unstable as a drug form to be used directly in cancer chemotherapy. Based on preliminary data, the hypothesis being evaluated in this proposal is that encapsulation of 5'-triphosphates of antiproliferative NA in a submicron polymeric carrier with protective and targeting properties will result in a novel therapeutic form of the old drugs. The proposed formulation and delivery system is based on self-assembled polyionic complexes formed between nucleoside 5'-triphosphates and cationic carrier called 'Nanogel'. This carrier consists of a cross-linked network of cationic polyethylenimine and poly (ethylene glycol) or Poloxamer block copolymers. Nanogel loaded with triphosphate nucleotides in aqueous media forms small nanosized particles. Formulated into the particles for systemic administration, active triphosphates of NA can be conventionally stored in freeze-dried form and then readily dispersed before injection. Nanogel can protect triphosphate nucleotides in circulation against enzymatic degradation and drastically increase intracellular transport of anionic nucleotides, which otherwise is not effective. Specific aims of the proposal are to: (1) formulate polymer-nucleotide complexes with increased dispersion stability and enzymatic resistance, (2) Determine whether the polymer-nucleotide complexes can increase the cytotoxic effects of nucleotide analogues, and (3) Examine how the polymer-nucleotide complexes can enhance the systemic therapy of tumors in vivo. A panel of representative NA and cancer cell lines will be studied, and a murine Lewis lung carcinoma model will be used to verify obtained in vitro results. The long circulating polymer-nucleotide complexes can display better tumor accumulation because of the 'enhanced permeability and retention' effect. They can also be modified by vector ligands with affinity to surface receptors on actively proliferating cancer cells in order to enhance selective accumulation of the cytotoxic NA in tumors or metastatic nodes. Application of the drug forms may help to prevent many of the known chemotherapy side effects. Data accumulated in these studies can be directly used for design of better systemic formulations of cytotoxic nucleotide drugs.

Award # 5R01CA119414-02 Agency NIH Lead Institution University of California-San FranciscoTypeExtramural

Explanatory Notes The goal of this grant is to develop hybrid gold-nanoparticle-based molecular imaging and therapeutic agents for diagnosis and treatment of prostate cancer.The EHS relevant component of this research includes characterization and study of photophysical properties, size, dispersity, biolocalization, pharmacokinetics and in vivo profiles of stabilized gold nanoparticles.

Abstract This project will develop targeted nano-probes for molecular imaging to enable non-invasive early detection of incipient cancer, affording substantive improvements in sensitivity and selectivity. It brings together three research groups with complementary expertise in angiogenesis and mouse models of cancer (Hanahan, UCSF), in vascular profiling (Ruoslahti, Burnham Institute), and in clinical and experimental molecular imaging (Franc, UCSF). Peptides have been discovered that specifically home through the circulatory system to the angiogenic blood or lymphatic neo-vasculature of high-grade neoplasias and/or invasive carcinomas. These vascular signatures can distinguish cancerous lesions from their cognate normal tissue, as well as from blood/lymphatic vessels in other organs and neoplastic conditions. The aims are: 1. Develop imaging nanoprobes for detecting the blood and lymphatic neo-vasculature in two mouse models of cancer that undergo stepwise progression to carcinoma, using validated signature-finding peptides as modular specificity elements linked to agents appropriate for imaging with SPECT, PET, or MRI. 2. Discover and characterize a repertoire of new signature-finding peptides for blood and lymphatic vasculature of cervical and pancreatic ductal neoplasias and cancer, and determine which identify analogous lesions in the cognate human diseases. 3. Competitively evaluate mouse/human signature-finding peptides (and mixtures thereof) from Aim 2 to identify the best at delivering imaging reporters to the aberrant blood and lymphatic vasculatures in the mouse models of cervical (as a prototype) and pancreatic ductal cancer (for its an unmet clinical need). 4. In partnership with Centers of Excellence in Cancer Nanotechnology, test nano-probes in the mouse models consisting of the best human/mouse signature-finding peptides linked to new imaging nanostructures being developed by those centers, to identify optimal candidate nanoprobes for clinical evaluation. The modular imaging nanoprobes to be developed, by delivering imaging agents to organ sites of tumor angiogenesis and lymphangiogenesis, hold promise to enable early detection of human cancer

Award # 5R01CA119412-02 Agency NIH Lead Institution University of Missouri-ColumbiaTypeExtramural

Explanatory Notes The EHS relevance of this research is development and implemention of techniques to detect and characterize genetic damage that may result from a variety of chemicals and materials in the environment, including carbon nanotubes.

Abstract The University of Missouri (MU) Cancer Nanotechnology Platform (CNP) is the culmination of long-standing interdisciplinary partnerships in departments within the School of Medicine, the College of Arts and Science, the College of Veterinary Medicine, the Ellis Fischel Cancer Center, the Missouri University Research Reactor and the College of Engineering. Our CNP's overarching goal focuses on developing hybrid gold nanoparticle-based molecular imaging agents and targeted therapeutic agents that are effective in the diagnosis and treatment of prostate cancer. The specific objectives of our CNP include the following: (i) Synthesize and use a library of gold nanoparticles (AuNPs) for conjugation with prostate tumor-specific bombesin peptides. Investigate the photophysical properties, dispersity, and size-dimension measurements on these gold nanoparticles. (ii) Investigate biolocalization, pharmacokinetics and in vivo profiles of AuNPs stabilized with starch, agarose, and arabinagalactan protein (gum Arabic) in pigs. We will optimize analytical protocols, using techniques of neutron activation analysis (NAA), atomic absorption and radiochemical analysis using Au-198 nanoparticulate tracer, for the accurate estimation of AuNPs/gold metal in tissues and biological fluids, (iii) Develop new theoretical models, computations and simulations for the interaction of AuNPs with cells, (iv) Investigate the utility of AuNPs and bombesin-conjugated hybrid AuNPs as image enhancers in computed tomographic (CT) and ultrasound (US) imaging of prostate tumors in SCID mice implanted with human prostate cancer xenografts and with experimental modeling, simulations and studies on anthropomorphic phantom models of prostate cancer, (v) Optimize production of nanoparticulate a-emitting Au-198/199 and their utility for designing and developing tumor-specific Au-198/199-nanoparticlelabeled bombesin peptides for prostate tumor therapy. These studies will validate that multiple atoms of fiemitting Au-198 isotope, characteristic of nanoparticulate gold, will deliver significantly higher therapeutic payload to tumor sites than any conventional therapy. The MU Nanoparticles Production Core Facility will serve as a research and production resource for the unhindered supply of AuNPs to all investigators within the CNP program. The CNP provides unique opportunities for professionals from diverse academic backgrounds to be involved in collaborative scientific interactions that further our interdisciplinary research, education and product development program. We will apply principles of nanoscience and nanotechnology to develop innovative molecular imaging and therapeutic approaches to combat prostate cancer

Award # 1R01ES015498-01 Agency NIH Lead Institution University of California-Los AngelesTypeExtramural

Explanatory Notes The goal of this project is to develop novel cytotoxic nucleotide drugs embedded in a nanoscale polymer carriers that help stabilize, protect and target the drug. The EHS relevant component of this research is to determine whether the polymer-nucleotide complexes can increase the cytotoxic effects of the non-encapsulated nucleotide analogues.

Abstract Data for performing a preliminary risk assessment of manufactured nanomaterials are just beginning to emerge. However, early studies of nanomaterial toxicity in aqueous media have tended to be more observational than mechanistic, and have often focused on a single, advanced stage of toxicity that could yield contradictory results. Moreover, the ability to generalize findings to other nanomaterials is limited by the lack of a rational basis for categorizing nanomaterials. Elucidating the mechanisms of toxicity for a given nanomaterial will provide a basis for classifying materials for regulatory purposes, postulating dose-response curves, screening potential risks, and prescribing strategies for risk management. The primary objective of this work is to elucidate the mechanism(s) by which manufactured nanoparticles may induce toxicity in vitro and in vivo. Specifically, this study will consider fullerene-based materials, comparing them with, (i) reference standards (TiO2 and carbon black); (ii) ultrafine particles obtained from an urban airshed (well characterized by in vitro toxicology studies); We will explore a methodology for rapidly screening potentially toxic nanoparticles based on their propensity to generate ROS. The principal hypothesis is that certain classes of nanoparticles such as fullerenes induce ROS production, cellular oxidative stress and cytotoxicity. Fullerenes are selected based on the relatively novel properties (e.g. strength.arid electron affinity) that make them attractive for commercialization. The investigators propose that oxidative stress induced by fullerene derivatives occurs in several stages (tiers), beginning with the induction of phase II antioxidant defenses at the lowest tier of oxidative stress (tier 1), followed by pro-inflammatory (tier 2) and mitochondrion-mediated cytotoxic effects (tier 3) as the level of oxidative stress increases. Particle size, shape, surface area, charge, and chemical composition are important physical variables that could determine their ROS-generating or scavenging properties. Rapid physicochemical determination of ROS production might provide a paradigm to assess the possible toxicity of nanomaterials that act via these mechanisms. Specific Aim 1 will characterize commercial nanoparticles and their derivatives in terms of particle size, shape, surface area, charge, aqueous solubility, propensity to aggregate, and their ability to catalyze or quench ROS production in vitro. Materials will also be characterized in model solutions containing naturally occurring organic matter, proteins and ions at levels similar to those present in natural waters. Aim 2 will determine whether various fullerenes can generate a hierarchical oxidative stress response in macrophages, bronchial epithelial cells, endothelial cells, neural cells and hepatocytes. This will be accomplished by comparing the effects of fullerenes and reference nanoparticles on, (i) phase II enzyme expression and activation of the heme oxygenase 1 (HO-1) promoter (tier 1); (ii) cytokine and chernokine expression as well as assays for MAP kinase activation (tier 2); (iii) mitochondria! perturbation and induction of cellular apoptosis (tier 3). These biological responses will be compared to the physicochemieal properties of nanomaterials elucidated in Aim 1. Aim 3 will perform in vivo imaging of the oxidative stress-sensitive HO-1 promoter linked to a luciferase reporter in transgenic mice. Organs and tissues showing increased luciferase activity will be investigated for histological evidence of inflammation and cytotoxicity. Aim 4 will compare the biologic responses elicited by each of the nano-scale particles with their ability to generate ROS abiotically, and test the hypothesis that ROS generation can be used to screen toxicity. By focusing on mechanisms of toxicity rather than outcomes alone, this work will provide the basis for classifying nanomaterials for regulatory purposes. Based on preliminary results presented in this proposal, we anticipate that ROS generation in solution and under UV radiation will be good predictors of nanoparticle toxicity and that ROS measurements can be adapted to screen nanomaterials. A broader assessment of nanomaterial toxicity in the context of the hierarchical oxidative stress response is likely to yield a more sensitive paradigm for toxicity testing, perhaps resolving inconsistencies reported in the literature

Award # CA-R*-NEU-7524-H Agency USDA (CSREES) Lead Institution University of California-RiversideTypeExtramural

Explanatory Notes

Abstract OBJECTIVES: The objective of this proposal is extend our research on chromosomal alterations induced by environmental and agricultural chemicals to detect and characterize genetic damage occurring in cells in vitro, in animal models and in exposed human populations. A series of studies will be conducted focusing on agents exerting damage through direct and indirect genotoxic mechanisms. 1) Initial studies will focus on the development and application of techniques to detect cellular and genetic alterations in human and animal cells. 2) The next phase of this research will be to apply these techniques to detect genetic damage induced by selected environmental and model agents, and to conduct mechanistic studies to understand the mechanisms responsible for the observed genotoxic effects. 3) In the last phase, we will continue and extend our efforts to apply these techniques to detect chromosomal alterations in cells obtained from humans exposed to environmental chemicals or individuals at elevated risk for developing cancer. APPROACH: A series of studies will be conducted using conventional and molecular cytogenetic approaches to identify genotoxic agents. The three phases are: 1) Initial studies will focus on the continued development and application of techniques to detect cellular and genetic alterations in human and animal cells. 2) The next phase of this research will be to apply these techniques to detect genetic damage induced by selected environmental and model agents, and to conduct mechanistic studies to understand the mechanisms responsible for the observed genotoxic effects. 3) In the last phase, we will continue and extend our efforts to apply these techniques to detect chromosomal alterations in cells obtained from humans exposed to environmental chemicals or individuals at elevated risk for developing cancer. We propose to extend our earlier studies to demonstrate the feasibility of utilizing fluorescence in situ hybridization and related techniques to detect chromosomal alterations in various rat and mouse tissues. We also plan to determine the feasibility measuring DNA damage-responding proteins within cells as a screen for detecting clastogenic and aneugenic agents. We proposed to build upon our in vitro work using proliferating human cells to explore the use this technique to detect chromosomal damage in resting (Go) cells in vitro, and in resting and dividing cells in rodents exposed to model and environmental agents in vivo. We propose to continue our in vitro and in vivo studies of benzene and o-phenylphenol (OPP), two widely used environmental carcinogens. We propose to perform these types of tests on other agents of current concern such as carbon nanotubes, propoxur metabolites, mosquito coil-releasing pesticides or related products. Studies specifically focusing on the mechanisms underlying the observed toxic and genotoxic effects will also be performed. For studies on benzene, we propose to continue our studies on the potential role of inhibition of topoisomerase II in the toxicity and clastogenicity of this agent. For OPP, we propose to investigate the role of a prostaglandin[H]synthase in the bioactivation of the OPP metabolite phenylhydroquinone in the rat bladder. We propose to continue our studies using FISH and conventional cytogenetic approaches to determine frequency of alterations in cells obtained from exposed human populations. These studies will be initiated as opportunities arise. Using the results from our previous studies, we propose to look for associations between polymorphisms in genes involved in xenobiotic metabolism and DNA repair and frequencies of chromosome breakage and aneuploidy. This should provide additional insights into individuals at risk for chromosomal damage as well as provide insights into the mechanisms underlying the genotoxic effects. As an extension of our biomonitoring studies, we also plan to continue our studies to determine if these FISH techniques can be used to identify individuals at elevated risk of developing cancer and improve cancer diagnosis. We believe that these more clinical studies will also provide mechanistic insights into the development of specific cancers.

Award # 1R43DK075190-01 Agency NIH Lead Institution Therapyx, IncTypeExtramural

Explanatory Notes This research seeks to optimize a system for non-invasive administration of medication encapsulated in nanoparticles. Dosage and kinetic studies are a major research thrust.

Abstract Type 2 diabetes mellitus is one of the leading causes of death and disability in the world. While current treatments prevent some of diabetes' more damaging complications, they can neither restore normoglycemia nor eliminate all the harmful effects caused by long term diabetes. Recent studies have shown that the continuous intravenous (i.v.) administration of Glucagon-like peptide 1 (GLP-1) can lower plasma glucose significantly in a majority of patients with Type 2 diabetes. However, due to the need for the continuous infusion of GLP-1 and the failure of oral GLP-1 peptide to lead to an increase in serum GLP-1 levels, current approaches using GLP-1 are not suitable for long term diabetes treatment. Advanced controlled release systems provide an alternative approach to the standard oral, intravenous or intramuscular delivery of pharmaceutical agents. This application proposes a non-invasive diabetes therapy through the oral administration of GLP-1 encapsulated in biodegradable nanoparticles. Encapsulation of labile biologicals such as GLP-1 into biodegradable polymers provides protection from the acidic environment and proteases of the stomach and allows safe passage into the lumen of the Gl tract for uptake. This proposal is designed to determine whether orally delivered, nano-encapsulated GLP-1 are superior to systemic bolus delivery in reducing blood glucose levels in experimental animals. To this end, three aims are proposed. In Aim 1, different bioadhesive formulations of encapsulated GLP-1 are analyzed for release kinetics (ELISA) and post-release peptide bioactivity (INS-1 cell insulin release / proliferation) in vitro. The goal of Aim 2 is to determine in vivo dosing and kinetics. In this aim, control mice are treated with a intra-peritoneal injection of glucose followed by the oral administration of GLP-1 nanoparticle formulations selected from the release and bioactivity studies in Aim 1. In the final Aim, the therapeutic efficacy of GLP-1 nanoparticles is examined in fasting and post-prandial diabetic mice. Type II diabetes mellitus and its long term complications affect 18.2 million people in the United States alone. Deaths due to the long-term complications of this disease make diabetes the third leading cause of death in the US. The orally delivered form of GLP-1 proposed here could have a profound effect on public health due to its potential to reduce the illness and death associated with long term diabetes

Award # 3U10HD037242-08S1 Agency NIH Lead Institution Baylor College of MedicineTypeExtramural

Explanatory Notes

Abstract The primary aim of this application for the Mentored Specialized Clinical Investigator Development Award (MSCIDA) in Pediatric Pharmacology is to provide a comprehensive training program in pediatric clinical pharmacology for Dr. Patrick A. Thompson, a board-eligible pediatrician and sub-specialty fellow in hematology-oncology. Patrick has a unique background in engineering, mathematical modeling and clinical medicine. This comprehensive training program for Dr. Thompson has been tailored to 1) further develop his capabilities in pharmacokinetic modeling and pharmacokinetic, pharmacodynamic and pharmacogenetic correlations, 2) provide him with the knowledge and ability to design, conduct, and analyze pediatric clinical therapeutics trials, 3) give him a thorough understanding of the complex regulatory issues associated with pediatric labeling of drugs and biology, and 4) teach him to carry out translational research in pharmacology. His training will occur through Baylor College of Medicine's (BCM) Pediatric Pharmacology Research Unit (PRRU) and will include formal enrollment in the BCM Clinical Pharmacology Fellowship Program, which is registered with the American Board of Clinical Pharmacology. The BCM Clinical Pharmacology fellowship program curriculum covers all the topics required by the MSCIDA award. It includes participation in BCM's, NIH-K30 grant-supported Clinical Scientist Training Program (CSTP), formal coursework at the University of Houston College of Pharmacy, and clinical rotations through the University of Texas and Texas Children's Hospital. The didactic component of this fellowship also includes outside rotations at the Food and Drug Administration, with the pharmaceutical industry, and with experts in population-based modeling at a collaborating PPRU site. The research component of Dr. Thompson's career plan includes two projects: 1) a pharmacometric project to evaluate the pharmacokinetic variability of an important anticancer agent, irinotecan, utilizing population modeling techniques; and 2) a project to characterize the pharmacology and biodistribution of two types of therapeutic antibody-targeted gold nanoparticles that have promising applications for the treatment of malignant or other disease processes. The proposed Career Development Plan for Dr. Thompson will provide an excellent foundation for achieving his long-term goal of becoming a leading clinician-scientist and educator in pediatric clinical pharmacology

Award # 5U10HD037261-07 Agency NIH Lead Institution Wayne State UniversityTypeExtramural

Explanatory Notes

Abstract The overarching aim of this proposal is to ensure continuation of the Neonatal and Pediatric Pharmacology Research Unit (PPRU) at the Children's Hospital of Michigan and Wayne State University (WSU) The WSU PPRU has conducted more than 40 multicenter and single site trials and has led the evaluation of drugs such as the multicenter tiral on the new formulation of intravenous ibuprofen for early closure of patent ductus artriousus in the preterm newborn. WSU PPRU ranked among the top 6 sites for subject enrollment. In addition, studies on aberrant development of serotonin biosynthesis in autistic children using PET scanning and identification of the phenotypic variants of serotonin metabolism have led to the ongoing evaluation of buspirone in drug therapy of autism. The demonstration of the role of NFKb in the gene expression of inflammatory mediators including the cyclo-oxygenases identified a target focus for drug intervention. The specific aims include: 1) the conduct of studies to generate requisite data on bioavailability, formulations, drug metabolism, pharmacokinetics, pharmacodynamics, safety, and effectiveness of new molecular entities and drugs currently used and are potentially used in the unborn fetus, newborn and children. 2) To determine the ontogeny of GABA receptors and to develop innovative pharmacotherapies of neonatal and childhood diseases such as neonatal apnea, neonatal hypoxic-ischemic encephalopathy and autism 3) To apply nanotechnology and use of dendrimers in targeted drug delivery and to develop novel formulations to modulate drug transfer across the blood brain barrier during development 4) to determine the validity of non-invasive cry analyses technique as surrogate pharmacodynamic measure in neonatal pain and non-verbal children 5) to develop novel drug therapies for fetuses with intrauterine growth retardation. 6)To determine the role of B2AR gene polymorphism in beta adrenergic response in asthmatic children and to provide a clinical milieu for the application of pharmacogenomics and proteomic technology leading to individualized pediatric pharmacotherapy and 7) to provide educational and training programs in pediatric pharmacology. Taken collectively, these initiatives will advance rational, safe, effective and cost beneficial drug therapies in the fetus, newborn and children

Award # 5S11ES013339-02 Agency NIH Lead Institution University of Texas-El PasoTypeExtramural

Explanatory Notes This work seeks to develop data relating ultrafine (less than 100 nanometer) air particulate contamination with asthma causation. Baseline information on the characteristics of ultrafine particles will be developed, including specific focus on occurrence of carbon nanotubes in kitchens and in vitro data on inflammatory measures following carbon nanotube exposure.

Abstract This revised application for an Advanced Research Cooperation in Environmental Health (ARCH) grant links the University of Texas at El Paso (UTEP), a Minority Serving Institution (MSI), with the University of New Mexico Health Sciences Center (UNM HSC), a Research Intensive University (RIU). The central research hypothesis is that children breathing air in the most polluted parts of El Paso, TX, have an increased prevalence of asthma, which may be under detected in the area's medically underserved Hispanic children. The project builds on the research strengths and previous work of UTEP investigators in documenting significant air and soil pollution problems in El Paso, TX, and contiguous Juarez, Mexico, and on UTEP's recent successful programs in public health. The Core Research Project ties high-density (both spatial and temporal) air and soil quality data to the prevalence and intensity of asthma and respiratory distress in a cohort of 1200 households randomly selected from 100 stratified blocks in the El Paso community. The application has been completely revised with an improved environmental sampling design, development of a more expansive cohort, new environmental epidemiology expertise, and more productive interactions of all Pilot Projects with the Core Research Project. The Pilot Projects will monitor levels of pollutants, including PM2.5, carbon nanoparticles, toxic metals, polycyclic aromatic hydrocarbons (PAHs), atmospheric ozone, nitrogen dioxide (NO2), and volatile organic compounds (VOCs), and feed these data to the Research Core. In revised Project 1, lung function in children measured by Impulse Oscillometry (IOS) methods in a clinical setting will be compared to lung function measured with spirometry methods promotoras in the local communities. Project 2 is new and examines the relationship between indoor and outdoor air sample particle concentration and composition in a subset of the blocks in the Research Core Project. Project 3 is also new and will test for gases (NO2, ozone, and VOCs), again in neighborhoods tied to the Core. Revised Project 4 examines the hypothesis that the organic fraction of PM2.5 contributes to asthma via oxidized PAHs that activate cell signaling pathways important in inflammation and immediate type hypersensitivity leading to asthma exacerbations. Project 5 performs innovative studies on carbon nanotubes from local environmental sources. The ARCH Program is overseen by an Administrative Core and is supported by a Facility Core at UTEP

Award # E2156_NCTR Agency NIH Lead Institution TypeExtramural

Explanatory Notes

Abstract National Toxicology Program

Award # Agency NIH Lead Institution TypeExtramural

Explanatory Notes

Abstract National Toxicology Program

Award # 1R01ES015498-01 Agency NIH Lead Institution University of California-Los AngelesTypeExtramural

Explanatory Notes

Abstract Data for performing a preliminary risk assessment of manufactured nanomaterials are just beginning to emerge. However, early studies of nanomaterial toxicity in aqueous media have tended to be more observational than mechanistic, and have often focused on a single, advanced stage of toxicity that could yield contradictory results. Moreover, the ability to generalize findings to other nanomaterials is limited by the lack of a rational basis for categorizing nanomaterials. Elucidating the mechanisms of toxicity for a given nanomaterial will provide a basis for classifying materials for regulatory purposes, postulating dose-response curves, screening potential risks, and prescribing strategies for risk management. The primary objective of this work is to elucidate the mechanism(s) by which manufactured nanoparticles may induce toxicity in vitro and in vivo. Specifically, this study will consider fullerene-based materials, comparing them with, (i) reference standards (TiO2 and carbon black); (ii) ultrafine particles obtained from an urban airshed (well characterized by in vitro toxicology studies); We will explore a methodology for rapidly screening potentially toxic nanoparticles based on their propensity to generate ROS. The principal hypothesis is that certain classes of nanoparticles such as fullerenes induce ROS production, cellular oxidative stress and cytotoxicity. Fullerenes are selected based on the relatively novel properties (e.g. strength.arid electron affinity) that make them attractive for commercialization. The investigators propose that oxidative stress induced by fullerene derivatives occurs in several stages (tiers), beginning with the induction of phase II antioxidant defenses at the lowest tier of oxidative stress (tier 1), followed by pro-inflammatory (tier 2) and mitochondrion-mediated cytotoxic effects (tier 3) as the level of oxidative stress increases. Particle size, shape, surface area, charge, and chemical composition are important physical variables that could determine their ROS-generating or scavenging properties. Rapid physicochemical determination of ROS production might provide a paradigm to assess the possible toxicity of nanomaterials that act via these mechanisms. Specific Aim 1 will characterize commercial nanoparticles and their derivatives in terms of particle size, shape, surface area, charge, aqueous solubility, propensity to aggregate, and their ability to catalyze or quench ROS production in vitro. Materials will also be characterized in model solutions containing naturally occurring organic matter, proteins and ions at levels similar to those present in natural waters. Aim 2 will determine whether various fullerenes can generate a hierarchical oxidative stress response in macrophages, bronchial epithelial cells, endothelial cells, neural cells and hepatocytes. This will be accomplished by comparing the effects of fullerenes and reference nanoparticles on, (i) phase II enzyme expression and activation of the heme oxygenase 1 (HO-1) promoter (tier 1); (ii) cytokine and chernokine expression as well as assays for MAP kinase activation (tier 2); (iii) mitochondria! perturbation and induction of cellular apoptosis (tier 3). These biological responses will be compared to the physicochemieal properties of nanomaterials elucidated in Aim 1. Aim 3 will perform in vivo imaging of the oxidative stress-sensitive HO-1 promoter linked to a luciferase reporter in transgenic mice. Organs and tissues showing increased luciferase activity will be investigated for histological evidence of inflammation and cytotoxicity. Aim 4 will compare the biologic responses elicited by each of the nano-scale particles with their ability to generate ROS abiotically, and test the hypothesis that ROS generation can be used to screen toxicity. By focusing on mechanisms of toxicity rather than outcomes alone, this work will provide the basis for classifying nanomaterials for regulatory purposes. Based on preliminary results presented in this proposal, we anticipate that ROS generation in solution and under UV radiation will be good predictors of nanoparticle toxicity and that ROS measurements can be adapted to screen nanomaterials. A broader assessment of nanomaterial toxicity in the context of the hierarchical oxidative stress response is likely to yield a more sensitive paradigm for toxicity testing, perhaps resolving inconsistencies reported in the literature

Award # Agency NIOSH Lead Institution NIOSH/HELDTypeIntramural

Explanatory Notes

Abstract With the potential mass production of nanomaterials, there is a need to determine if health effects from occupational exposures to these materials may result. Precious studies heave indicated that inhalation exposure to small particles can exacerbate pre-existing conditions such as asthma, COPD and cardiovascular disease. Current studies are designed to address whether carbon nanotubes can influence lung inflammatory mediators, COPD and cardiovascular functions employing conventional and transgenic animal models. These studies will determine whether nanoparticles, and which types of nanoparticles, have the potential to influence lung diseases, including asthma, fibrosis and COPD as well as coronary artery disease in exposed workers. Results from these studies should provide important information on hazard identification and dose-response which can be used in determining risk in the workplace

Award # Agency NIOSH Lead Institution NIOSH/HELDTypeIntramural

Explanatory Notes

Abstract Nanoparticles are new materials of emerging technological importance in different industries. Because dermal exposure is likely in a number of occupational settings, it is very important to assess whether nanoparticles could cause adverse effects to skin. The hypothesis is that nanoparticles are toxic to the skin and the toxicity is dependent on their penetration to skin, induction of oxidative stress, and content of transition metals. Because inflammation provides a redox environment in which transition metals can fully realize their pro-oxidant potential, a combination of inflammatory response with metal oxide particles, or iron-containing SWCNT will synergistically enhance damage to cells and tissue. Results obtained from these studies provide critical knowledge about mechanisms of dermal toxicity of nanoscale materials and will be used by regulatory agencies (OSHA and EPA) and industry to address strategies for assurance of healthy work places and a safe environments.

Award # Agency NIOSH Lead Institution NIOSH/HELDTypeIntramural

Explanatory Notes

Abstract Recent years have seen an exponential growth in the development and production of nanomaterials. These materials have unique physical, chemical and electrical properties due to specially forged arrangements of atoms on a nanometer scale that do not occur in natural systems. Because of the unique properties and small size of nanoparticles, issues have been raised as to their potential adverse effects on the lung upon inhalation and whether they can translocate to systemic sites. This project will identify where in the lungs inhaled nanomaterials might deposit, the health risks that might arise from nanomaterial deposition, and to what extent the nanomaterials might translocate to other organs of the body after depositing in the lungs. Results of this study will address critical issues identified by the NIOSH Nanotechnology Research Center and assist in hazard identification and risk assessment.

Award # 0540920 Agency NSF Lead Institution University of FloridaTypeExtramural

Explanatory Notes

Abstract The proposal is concerned with biological, histological and molecular studies of toxic effects of metal nanoparticles on zebrafish. The fish will be exposed for 96 hours to aqueous suspensions of aluminum, silver, nickel, and titania nanoparticles of various concentrations. Toxicity effects will be evaluated at the morphological, biochemical and molecular levels in gills, liver, intestine, and kidney tissues. One of the main objectives is to compare toxicity of metals in the dissolved and nanoparticle state as well as the nanoparticles of different sizes and shape. The hypothesis is that the metal nanoparticles will produce toxic effects that are different from that of metals in aqueous solution. The proposal will take advantage of newly arriving zebrafish microarrays to characterize gene expression at different treatment levels, followed by quantitative PCR of the 15 most prominent genes. An in vitro model system will also be tested as an alternative for a toxicity assay of metal nanoparticles in aquatic species.

Award # Agency DOD (AFOSR) Lead Institution TypeIntramural

Explanatory Notes

Abstract Thanks to in-house funding, AFRL scientists are accomplishing a multitude of nanotechnology research projects geared towards investigating the biological
interactions of engineered nanomaterials, including potential toxicities arising from the physicochemical properties uniquely associated with nanoscale structures. This research will acquire the fundamental knowledge to facilitate a better understanding of nano-bio interaction mechanisms, provide in-depth analyses of corresponding effects on biological systems and enable the theoretical development of predictive bioresponse models. Such knowledge will not only help to improve nanomaterial safety strategies for the protection of both human and environmental health, but also help to apply advanced nanobiotechnologies to the development of future weapon systems.

Under certain conditions of exposure, every chemical is toxic. While the AF is working to minimize issues concerning the production, handling, and disposal of nanomaterials as they relate to future mission requirements, a significant knowledge gap remains with respect to the human and environmental health implications of increasing nanomaterial usage. There is a need to understand the potential sources and effects of exposure to nanomaterials throughout their cycles of production and use. Further, it is critical to understand the transport—and evolution—of nanomaterials channeled through the environment and the human body, including their significant avenues of access and potentially adverse effects.

Two key areas of immediate military relevance include propulsion and munitions systems, which employ nanomaterials not only in tuning systems for greater insensitivity to ignition during storage and delivery, but in generating more energetic propulsion/explosions and ensuring long-term storage stability. Consequently, there is a growing need for nanoenergetics and other novel technologies to satisfy the increasing performance demands of propulsion and munitions systems.

Award # Agency DOD (AFOSR) Lead Institution ONAMITypeExtramural

Explanatory Notes

Abstract The scope of the work involves the continuing development of the Safer Nanomaterials and Nanomanufacturing Initiative, a research collaborative within the Oregon Nanoscience and Microtechnologies Institute (ONAMI). The Initiative shall use proactive strategies to develop new nanomaterials and nanomanufacturing approaches that offer a high level of performance, yet pose minimal harm to human health or the environment. The work will merge the principles of green chemistry and nanoscience to produce safer nanomaterials and more efficient nanomanufacturing processes in the context of producing nanoparticles and nanostructured materials applications in photovoltaics,nano-electronics and sensing. This work represents a collaborative effort that includes researchers from the University of Oregon (UO), Oregon State University (OSU), Portland State University (PSU), and Battelle - Pacific Northwest National Laboratory (PNNL) and is divided into three broad thrust groups: 1. designing safer nanomaterials (divided into Tasks 1-3); 2. greener nanomanufacturing of engineered nanoparticles (divided into Tasks 4-6) and; 2. interfacing nanoparticles to nano- and micro structures for device applications (divided into Tasks 7-9).

Award # CA-R*-NEU-7524-H Agency DOD (AFOSR) Lead Institution University of California-RiversideTypeExtramural

Explanatory Notes The EHS portion of the goal of this grant is to determine the relationship between the toxicity and physicochemical characteristics of nanoparticle by developing methodologies to characterize nanoparticles and test nanomaterials for toxicity using in vitro and in vivo techniques.

Abstract Nanotechnology present new opportunities to increase the performance of traditional products and to develop unique ones. Nanoparticle applications are already becoming more prominent in coatings, computers, clothing, cosmetics and other products and the trends suggest that nanoparticles will permeate a variety of industries. To ensure that this revolution continues, industry must guarantee that nanomaterials do not adversely affect human health but there is currently no systematically methodologies developed to characterize and test nanomaterials. Our goal is to determine the relationship between toxicity and physicochemical characteristics
(i.e. shape, size, surface chemistry, etc.) of nanoparticles and establish model systems as well as a battery of screening assays to determine the toxicity of different nanoparticles. In phase I, we propose to test two physicochemical properties –size and hydrophobic/hydrophilic coatings – to determine cytoxicity, inflammation (by assessing cytokine production) and glutathione levels as a marker of oxidative stress in a complex in vitro lung model system. In Phase II, we will expand the type of nanoparticles as well as the physico-chemical characteristics tested. In addition, we will establish an in vitro skin model and for those nanoparticles that elicited toxicity in our in vitro models an in vivo model to determine nanoparticle translocation.

Award # R833336 Agency EPA Lead Institution University of UtahTypeExtramural

Explanatory Notes

Abstract Objective: We will test the hypothesis that ingested nanoparticles are taken up by inflamed colon cells, translocate to the nucleus, and cause alteration of gene transcription. Colon cancer is a major cause of death, and any environmental agent that causes increased colon inflammation or affects the regulation of cell cycle genes is a logical concern. The goal of this toxicology study will be to quantify the ability of a suite of manufactured nanoparticles to affect inflamed versus non-inflamed colon cells and to elucidate the biochemical mechanisms linking particles to changes in gene regulation. This study is motivated by drug delivery studies showing that inflamed colon cells internalize nano particles, a paper reporting that silica particles translocate and bind nuclear proteins, and our preliminary gene array data showing that nano-sized silica affects the transcription of many genes associated with inflammation and cell cycle regulation. The research will focus on lower-cost carbon and metal oxide nanomaterials that are used commercially and therefore present the potential for human exposure by ingestion. Approach: The research will use established cellular imaging, molecular biology, and gene array technology and will build on the investigators' existing projects in particle toxicology, cell signaling, and colon cancer. Particles will be characterized using electron microsopy, surface area analysis, and trace element analysis. An in vitro model consisting of cultured human colon-derived cells (CaCo-2 and RKO cells) will be treated with suspensions containing the nanoparticles or controls. Tissue inflammation will be modeled by pretreating . Particle uptake and intracellular localization will be the cells with TNF- quantified by trace element analysis and by fluorescent probes. Particle-induced pro- and anti-inflammatory response will be measured by cytokine ELISA assays and cell death assessed by flow cytometry. Effects of the particles on gene regulation will be determined at the Microarray Core Facility using Agilent Full Human Genome 44K arrays, with confirmation of findings by quantitative real-time PCR. Expected Results: This exploratory study will test whether ingestion of certain commercially available manufactured nanoparticles is associated with altered inflammation and cell cycle pathway gene regulation in colon cells. The results will be useful for establishing whether the hypothesized effects represent a potential risk to human health and will help determine whether further animal or human exposure studies are justified.

Award # 5R21CA112436-02 Agency NIH Lead Institution Case Western Reserve UniversityTypeExtramural

Explanatory Notes This project tests nanomaterials in in vitro binding study and cell uptake study

Abstract Targeted drug delivery via nanoparticles holds great potential for the successful treatment of many deadly diseases such as cancer. PEG-modification is commonly used to prevent nonspecific interactions of the nanoparticles with blood components and nontarget cells. However the ability to achieve high targeting efficiency at the tumor site remains a significant challenge. There are several reports showing a reduction of transfection efficiency and decreased adhesion of phospholipids, liposomes and polymersomes presumably due to shielding of the targeting functional groups by the PEG chains. PEG chain length, grafting density, nanoparticle size and density of functional groups all were found to strongly influence the success of targeting. To analyze the influence of all these factors experimentally is a very complicated and time-consuming task. Computer simulation may considerably aid in this task by analyzing the influence of multiple parameters, thereby guiding the experimental research. The objective of the present proposal is to apply a combined theoretical and experimental approach to gain fundamental understanding of the physical principles of targeting by complex polymer nanoparticles. The central hypothesis is that the ligand valence and the structure of the corona of nanoparticles are the key factors defining the efficiency of site-specific targeting. To test this hypothesis we will carry out research designed to fulfill the following aims: 1) Structural design of a nanoparticle for effective targeting for each of the following concepts: a) using multivalent ligands and b) using short non-functional and long functionalized PEG chains in nanoparticle corona. 2) Design and experimental testing of the optimal PCL-PEG-cRGD nanoparticles for doxorubicih (DOX) delivery. Nanoparticle designs found to be optimal in simulations will be tested experimentally using in vitro binding study and cell uptake study in integrin-expressing and non-expressing melanoma cells. Successful execution of this research will supply an experimental practitioner with specific recommendations concerning the grafting density, PEG chain length, architecture, nanoparticle size and density of functional groups to ensure effective targeting of drug-containing nanoparticles. Because of the exploratory nature of the research some new approaches to drug targeting may be discovered. This knowledge will greatly facilitate the development and implementation of a new generation of drug delivery vehicles for cancer therapy

Award # 5R01CA112075-03 Agency NIH Lead Institution Scripps Research InstituteTypeExtramural

Explanatory Notes The EHS component investigates the behavios of viral nanoparticles in vivo.

Abstract Nanotechnology holds great promise for the early detection and treatment of cancer. The ability to both detect and follow the initiation and progression of cancer without biopsy, surgery, or other invasive techniques should lead to lower costs and higher quality of life. The goal of this proposal is to develop viral nanoparticles (VNPs) as platforms for combination tumor targeting and imaging agents in vivo. Our multidisciplinary team combines molecular biology, structure, chemistry and in vivo biology to attack this problem. In this proposal we will study two well-characterized viruses: a plant virus, cowpea mosaic virus (CPMV); and an insect virus, flockhouse virus (FHV). The accumulated knowledge of the structure, function, assembly, genome packaging, chemical attachment, and in vivo bioavailability of FHV and CPMV, developed in the co-investigators' laboratories, makes these viruses ideal candidates for such platform development. We will test the ability of tumor-specific VNPs to attack tumor cells in vitro and in vivo, and study mechanisms of uptake of VNPs into cells. We will also package anti-tumor compounds and inhibitory genes inside the particles to facilitate tumor destruction. Finally we will test the ability of our VNPs to detect and destroy tumors in vivo. These studies will make a significant contribution to the development of improved tumor targeting and imaging nanotechnologies

Award # 5R01CA107268-04 Agency NIH Lead Institution University of California-San FranciscoTypeExtramural

Explanatory Notes The goal of this grant is to develop non-viral gene nanoparticle carriers to target brain tumors. The EHS-relevant component of this research is to characterize the factors that optimize the ability of nanolipoparticles to target brain tumors and enhance the therapeutic efficacy of nanolipoparticles using in vitro and in vivo techniques.  

Abstract The objectives of this research are to devise improved non-viral gene carriers and to apply them in a Gene-Directed Enzyme Prodrug Therapy (GDEPT) protocol in a rat brain human tumor model. The question we will address is: will an optimized non-viral GDEPT protocol expressing yeast cytosine deaminase provide better therapy then a targeted liposome chemotherapy protocol in brain tumors? The hypothesis guiding this plan is that a combination of multiple levels of targeting will result in robust tumor-specific cytosine deaminase gene expression and improved tumor therapy. The four levels of targeting employed in these plans are: convective enhanced diffusion (CED) into the brain tumor, bioresponsive liposome gene formulations, CD44 receptor targeting, and a tumor matrix targeted enzyme. Tumor-localized cytosine deaminase will then convert the systemically administered prodrug, fluorocytosine (FC), to fluorouracil (FU), the active agent. This particular combination of approaches will enhance access of the carrier to the tumor, improve percolation of the carrier through the tumor, increase carrier attachment to tumor cells and improve intracellular delivery of the plasmid and bystander activity of the therapeutic gene. There are four major activities in the program: (1) synthesis of novel components and assembly of the components with plasmid DNA into a small diameter, stable liposome designated a nanolipoparticle (NLP)(2) characterization of the factors that control the performance of the NLP in cells (pH sensitivity, cell association, gene transfer activity, toxicity) and in animals (distribution, pharmacokinetics, toxicity and gene expression) after CED into the brain. (3) Construction of an extracellular matrix targeted cytosine deaminase to improve the bystander effect. (4) Comparison of the therapeutic effect of the optimized GDEPT to the effect of fluoroorotic acid delivered in a CD44 targeted liposome of similar physicochemical characteristics as the NLP. Fluorocytosine and fluoroorotic acid both exert their therapeutic effect through FU. The concentration of FU and metabolites will be measured in the tumor and plasma. We w