California NanoSystems Institute

The unique properties of nanoscale materials provide exciting new opportunities in disparate fields ranging from computer science to medicine. However, while nanoparticles may be designed for a wide array of specific functions, each will likely interact with biological systems. Such bio-nano interface may be intentional, as is the case for pharmaceutical or cosmetic applications, or accidental, as the result of industrial pollution. Thus, as the production and diversity of nanoparticles increases, it will become increasingly important to understand the principles governing how nanoparticles interact with biological systems. Here, the capabilities of high throughput screening (HTS), initially developed to identify novel pharmaceutical drugs, will be discussed with respect to identifying principles governing the outcome of bio-nanomaterial interactions.

Dr. Bradley is interested in i) identifying host proteins that are usurped by bacterial pathogens and ii) understanding how these interactions promote virulence. Specifically, we are studying protein exotoxins produced by Bacillus anthracis (the causative agent of anthrax), Campylobacter jejuni (the most common cause of bacterial diarrhea in the U.S.), E. coli (a common agent of food poisoning), and related bacteria. To better understand these interactions, Dr. Bradley's lab utilizes a number approaches including i) proteomics, ii) chemical genetics and iii) somatic cell genetics. The Bradley lab is also interested in developing therapeutics and detection systems for bacterial pathogens based on nanotechnology and is collaborating with a number of labs at the CNSI.

Dr. Bradley received a B.A. in Biochemistry and Molecular Biology from U.C. Santa Cruz and a Ph.D. in Microbiology and Molecular Genetics from Harvard Medical School. He is currently an Assistant Professor in the department of Microbiology, Immunology & Molecular Genetics at UCLA and a member of the California NanoSystems Institute. In 2005, Dr. Bradley assumed the role of Director of the Molecular Screening Shared Resource, a high-throughput screening facility that provides robotic automation for biological and chemical screening.

Recent and ongoing work in our lab on the interaction of well characterized single-walled and multi-walled carbon nanotubes with bacterial (E. coli) cells will be presented. Direct contact of bacterial cells with the carbon nanotubes results in significant cell damage and loss of viability, as verified by several independent techniques. Single-walled carbon nanotubes exhibit a much stronger antibacterial activity than multi-walled carbon nanotubes. Possible mechanisms for the inactivation of the bacterial cells upon contact with the carbon nanotubes are discussed. The results point out to the potential biotoxicity of carbon nanotubes in aquatic environments. The finding may also be useful in the application of carbon nanotubes as building blocks for antimicrobial materials.

Menachem Elimelech is the Roberto Goizueta Professor and Director of the Environmental Engineering Program at Yale University. Professor Elimelech received his B.S. and M.S. degrees from the Hebrew University in Jerusalem and his Ph.D. from Johns Hopkins University in 1989 in Environmental Engineering. His research interests include (1) transport and adhesion of microbial pathogens, (2) membrane separations for desalination and water quality control, (3) processes involving nanoparticles and biomacromolecules, and (4) water, sanitation, and health in developing countries. Professor Elimelech was elected to the National Academy of Engineering in 2006 and was awarded the Athalie Richardson Irvine Clarke Prize in 2005.

There are diverse ways that exposure to nanoparticles may be harmful to organisms or cells. We briefly survey aspects of nanoparticle reactivity and stability that are important components of their toxicity. Nanomaterials composed of redox-active elements, including iron, manganese and cerium, are particularly reactive and can provoke potentially damaging chemical transformations. For example, magnetite nanoparticles possess the capability for the reductive dechlorination of organic pollutants, while metallic iron particles can stimulate the degradation of organic molecules by both reductive and oxidative pathways. Moreover, even chemically inert nanoparticles, or molecule-nanoparticle assemblies, may be activated by light absorption. In certain environments, photochemical processes involving nanoparticles are known to effectively generate reactive oxygen species. Inorganic nanoparticles are found in many aqueous environments, and are frequently formed by microbial activity, thus providing insight into cell-nanoparticle interactions. For example, sulfate reducing bacteria that precipitate zinc sulfide (ZnS) nanoparticles as a by-product of their metabolism also possess the ability to sequester this product extracellularly. It is proposed that this may be an evolved mechanism to avoid zinc or ZnS nanoparticle toxicity.

Benjamin Gilbert obtained a B.A. in Natural Sciences from Cambridge University in 1994 and a PhD from the École Polytechnique Fédérale de Lausanne in 2000. His graduate research incorporated synchrotron x-ray studies at the Synchrotron Radiation Center of the University of Wisconsin - Madison, for which he received the SRC Aladdin Lamp Award. Following post-doctoral research at UW - Madison and the University of California at Berkeley, he joined Lawrence Berkeley National Lab. in 2004. In April 2007 he was promoted to a career scientist position. Gilbert has made important contributions to the rapidly evolving field of nanogeoscience - the study of the properties and geochemical interactions of natural nanoscale minerals. He retains an interest in applying and developing synchrotron x-ray experiments and analysis methods for the study of complex biological and geological systems. Research accomplishments include: the discovery of stable cluster formation by iron oxyhydroxide nanoparticles; the development of a theoretical basis for experimental observations of a thermodynamically stable nanophase material; observation of structural transformations in ZnS nanoparticles associated with water binding; the identification of nanoscale silicate inclusions in zircons; and combined experimental and theoretical descriptions of the crystal chemistry of manganese oxides. These are reported in more than 40 peer-reviewed publications that include collaborations with scientists from many disciplines.

Functional nanocomposites for optical, microelectronic and medical device applications have been developed using a nanoparticle platform technology based on the laser pyrolysis method. This laser-driven chemical process is found to produce small, uniform, multi-elemental, and dispersible nanoparticles. The major technical challenges in the industrial applications have been (a) synthesis of high-quality nanoparticles with precisely-controlled surface properties, (b) superior dispersion of nanoparticles in either aqueous or organic solution, (c) surface modification over individual nanoparticles, and (d) linkage of nanoparticles to a polymeric material. Many aspects in these sequential processes can be commonly shared with nano-bio interface studies. Structural properties such as particle size or degree of nanoparticle agglomeration, and surface chemistry and physics are particularly focused to study organic molecule - inorganic nanoparticle interfaces. This talk provides an overview of laser-made nanoparticles and their interfaces with organic molecules in case of transition metal oxide, metal, and semiconductor nanoparticles. Biocompatibility of nanoparticles at a cellular level is discussed as an extension from that at a molecular level.

Dr. Kambe's extensive background in technology development and commercialization derives from his work as a research manager in semiconductor materials and devices at Nippon Telephone & Telegraph (NTT) and as a managing director at the International Center for Materials Research in Japan. He was the founder of ICMR-USA and is a founder of NanoGram Corporation. Dr. Kambe has published numerous papers and articles on applications of nanomaterials and nano-polymer composites. He holds a PhD in Electrical Engineering from the Massachusetts Institute of Technology and BS and MS degrees in Instrumentation Engineering from Keio University in Japan.

The public's interest in nanotechnology generally starts with a broad concept of "novel, unique, surprising" phenomena observed at the nanoscale, but then collapses to health and safety concerns about nanoscale particles. Manufacturers of fine particles, especially those with an extensive commercial history, find themselves utilizing historically acceptable descriptive terms, e.g. ultrafine particle or Ostwald ripening, in a time period where nanoparticle and nanotoxicology have greater currency. Colleagues in the fields of toxicology, especially those working in lung inhalation, face a similar challenge between the historical studies, explanations, "modes of toxicity" and the current concern about potential consumer exposure during a nanomaterial's life cycle. AEROXIDE® P25 (a fine particle exhibiting features and porosity at the nanoscale and hence termed by some as nanostructured titania) has been well studied in both fields. The presentation will provide an overview of the shape, size, surface properties, use, and physicochemical characterization of this material with commentary on statements made in the toxicological literature about AEROXIDE® P25.

Fred Klaessig is currently the Business Director of the Aerosil Business Line at Degussa Corporation, a subsidiary of Degussa AG. For the past ten years, he was the Technical Director for the Aerosil & Silanes Business Unit, where his responsibilities ranged from technical service to new product introduction to liaison with the R&D Department in Germany to regulatory matters. AEROSIL® is a trade name for fumed silica, which the company has manufactured for 60 years and which is often cited as an example of a nanoparticle. Fumed silica, fumed titania and other fumed metal oxides are utilized in many fields for reinforcement, rheology control, abrasion and UV absorption. In the recent years, the great interest in nanotechnology has raised safety and registration concerns about materials of this class. These issues, both everyday technology and EHS concerns, has led to greater involvement in ASTM (E56), ISO (TC229) and industry organizations focusing on this subject area.

Fred Klaessig has a B.Sc. in Chemistry from the University of California, Berkeley and a Ph. D. in Physical Chemistry from Rensselaer Polytechnic Institute. His industrial experience has been with Bio Rad Laboratories as a Quality Control Chemist and various management positions at Betz Laboratories, now a division of GE Water Services, where his responsibilities involved scale and corrosion control in many chemical industrial processes.

Because of the large number of new nanomaterials that are being produced, it is of increasing importance to develop a platform for safety and risk assessment. It is probably not advisable to follow the example of chemical industry where the production of more than 80,000 industrial chemicals has overwhelmed toxicological screening capabilities. Toxicity testing has only been achieved for a few hundred chemicals and as a result, new examples of chemical toxicity revealed every year, often with devastating consequences to humans and the environment. One of the principal stumbling blocks in assessing chemical toxicity has been the cost and the logistics to perform animal and in vivo studies. An intuitively more enlightened approach would be to develop high throughput screening methods that incorporate a limited number of toxicological injury mechanisms that can be related to the physicochemical properties of nanomaterials. I will discuss the emerging paradigms of toxicity that can be linked to the physicochemical properties of engineered nanoparticles with a view to outlining scientific principles that originate at the nano/bio interface and which determines whether the particle interactions are injurious or biocompatible. The major toxicological paradigm that have emerged in nanoparticle toxicity relates to the semiconductor, electronic, UV activation, and redox cycling chemistry of the particles, which allows them to induce tissue damage through the generation of oxygen radicals, electron-hole pairs and oxidant injury. It is possible to follow the oxygen radical generation and oxidant stress injury by abiotic methods as well as a set of hierarchical cellular responses that determine protective, pro-inflammatory, mitochondrial damaging and pro-apoptotic outcomes. An oxidant injury pathway could translate into adaptive, pro-inflammatory or pro-apoptotic cellular effects in the lung, cardiovascular system and the brain. Another important paradigm relates to the ability of nanoparticles to absorb circulatory or cellular proteins as a function of particle size, surface area, functionalized surface groups, charge, hydrophobicity/hydrophilicity etc. This could induce protein unfolding, fibrillation, thiol crosslinking and loss of function, which could lead to neurotoxicity, loss of enzymatic activity, and generation of immunological responses. The thermodynamic properties and free surface energy of nanoparticles as a function of particle size, composition, phase and crystallinity could be responsible for particle dissolution in a biological environment, leading to the generation of cytotoxicity through the release of toxic ions or chemicals. Data are also emerging that indicate that cationic nanoparticles exert toxicity through the so-called proton sponge hypothesis, which postulates that particle uptake via acidifying endosomes leads to cellular toxicity through endosomal rupture, cytosolic deposition and mitochondrial targeting. The particle size, state of aggregation/dispersion and hydrophobicity also plays an important role in determining cellular uptake, subcellular localization and targeting of subcellular organelles. I will demonstrate that it is possible to devise high throughput screening methods to capture each of these toxicological mechanisms, which can then be used to classify nanoparticles into potentially hazardous and potentially safe. If used as a preliminary screen for newly emerging nanomaterials, these predictive science-based approaches can help to determine which materials should undergo priority testing in animal and in vivo exposure models. The knowledge gained from this approach will also reveal which nanomaterial properties are useful to promote biocompatibility.

André Nel is Professor and Chief of the Division of NanoMedicine at UCLA. He runs the Cellular Immunology Activation Laboratory in the Johnson Cancer Center and the Laboratory for Nanosafety Research and Testing in the California NanoSystems Institute (CNSI) at UCLA. Dr. Nel's chief research interests are: (i) Nanomedicine and Nanobiology, including nanomaterial properties that may assist nanomaterial safety testing; (ii) The role of air pollutants in asthma, with particular emphasis on the role of oxidative stress in the generation of airway inflammation and airway hyperreactivity. These studies are funded by personal RO1 grants from the NIH, the NIAD-funded Asthma and Immunology Disease Clinical Research Center, an EPA STAR award, and a UC Lead Campus Program for Nanotoxicology Research. Dr Nel is the Principal Investigator of the UCLA Asthma and Immunology Disease Center, Co-Director of the Southern California Particle Center and Director of the UC Lead Campus Program for Nanotoxicology Research and Training. Dr. Nel obtained his M.B., Ch.B. (MD) and Doctorate of Medicine (PhD equivalent) degrees from the University of Stellenbosch in Cape Town, South Africa, and subsequently did Clinical Immunology and Allergy training at UCLA. Dr Nel served a Chair of a study section at the NIAID and is Chair of the Air Pollution Committee in the AAAAI. Dr. Nel is a member of the ASCI, AAAAI, AAI and the Western Association of Physicians.

Monolayer-protected nanoparticles provide effective vectors for the delivery of drugs and biomolecules. The ability to attach targeting functionality presents a means of delivering these carriers to target organs and tissues. Additionally, the capability of tuning surface properties provides systems that are rapidly internalized by cells, with intracellular targeting possible through the use of appropriate functionality.

In addition to their ability to be functionalized in modular fashion, gold nanoparticles provide a tunable method for payload release. Glutathione (GSH) is present in low micromolar concentrations extracellularly. Intracellular GSH concentrations, however, range from 1-10 millimolar. This dramatic increase in GSH concentration within the cell provides a novel means for release: GSH can displace thiol functionality from the particle surface. This displacement process can be used directly to release drugs and prodrugs. Moreover, the addition of anionic GSH can be used to change the surface potential of cationic particles, providing an effective means for release of electrostatically-bound DNA and proteins. All of these release processes are tunable via control of the monolayer structure, with release rate correlating with monolayer length.

Charles A. Goessmann Professor of Chemistry Department of Chemistry and Program in Molecular and Cellular Biology University of Massachusetts at Amherst Vince Rotello received his B.S. from Illinois Institute of Technology in 1985. He obtained his Ph.D. in 1990 from Yale University with Harry Wasserman in the area of natural products synthesis. From 1990-93, he was an NSF postdoctoral fellow with Julius Rebek Jr. at M.I.T. in the area of host-guest chemistry. Since 1993, Professor Rotello has been at the University of Massachusetts at Amherst as an Assistant Professor from 1993-1998, Associate Professor (1998-2001), Professor (2001-2005) and Charles A Goessmann Professor of Chemistry (2005-), with appointments in Polymer Science and Engineering, and the Program in Molecular and Cellular Biology. He has been the recipient of the NSF CAREER, and Cottrell Scholar award, as well as the Camille Dreyfus Teacher-Scholar, and the Sloan Fellowships. His research program spans the areas of devices, polymers, nanotechnology, and biological systems, with over 235 papers published to date. Most recently, Vince's research has explored the use of nanoparticles in biology, including protein and DNA surface recognition as well as drug and DNA delivery.

With the explosion of our knowledge in protoeconomics and systems biology and the advances in micro fluidics, nano optics, and computer science, we stand on the verge of the most exciting paradigm change in medicine.

The presentation will cover scientific insights from multi disciplinary fields of physiology, molecular biology, organic chemistry, and nanotechnology. And will demonstrate how these interactions of these multi disciplines has resulted in the first FDA approved biologically interactive chemotherapy. Personalized medicine and targeted therapy will be the standard of care and the development strategy for biopharmaceutical companies of the future.

Patrick Soon-Shiong, M.D., M.D., is the Founder, Chairman and Chief Executive Officer of Abraxis bioscience, Inc. He has devoted his career to developing next-generation technology to treat patients with life-threatening diseases. Dr. Soon-Shiong performed the first encapsulated islet transplant in a diabetic patient and developed the nanoparticle albumin-bound (nab^TM) technology platform. Dr. Soon-Shiong's research has been recognized by noted organizations with numerous national and international awards such as the Association for Academic Surgery Award for Research, the American College of Surgeons Schering Scholar, the Royal College Physicians and Surgeons Research Award, the Peter Kiewit Distinguished Membership in Medicine Award, and the International J.W. Hyatt Award for Service to Mankind. Dr. Soon-Shiong received the 2006 Gilda Club Award for the advancement of cancer medicine and is a recipient of a 2007 Ellis Island Medal of Honor. He is a co-inventor of over 50 issued U.S. patents and has published more than 100 scientific papers. Dr. Soon-Shiong serves on the Board of Directors for the National Institute of Transplantation, the Technology Council for the Center for Cancer Nanotechnology Excellence, two RAND advisory boards, and the Board of Trustees for the Saint John's Health Center.

Mesoporous silica nanoparticles (MSNs) have recently emerged as a promising type of nanomaterials for delivering small organic compounds such as anticancer drugs. In addition, delivery of nucleic acids into plant cells as well as delivery of small proteins such as cytochrome c by using MSNs have been reported. These materials are biocompatible and are inert. In collaboration with Drs. Jeffrey Zink and Andre Nel, we have examined cellular uptake of MSNs having a diameter of 130 nm and containing pores with 2 nm diameter. Fluorescent dye was attached inside the pores and the surface was modified with phosphonate groups to avoid aggregation. We have shown that these nanoparticles are taken up by a variety of human cancer cell lines. Use of inhibitors showed that the uptake involves an energy-dependent endocytosis mechanism resulting in lysosomal localization. We have also shown that MSNs can trap hydrophobic anticancer drugs such as camptothecin and deliver them to human cancer cells causing cell killing. Finally, cancer targeting ligands attached to the nanoparticles dramatically increased the uptake of MSNs into cancer cells. These results point to a number of properties of the mesoporous silica nanoparticles that are favorable for their use as a delivery vehicle.

Fuyu Tamanoi is a biochemist who has served on the UCLA School of Medicine and UCLA College faculty since he joined the Department of Microbiology, Immunology & Molecular Genetics in 1993. He became a full professor in 1997. Since 1996, he has been a Director of Signal Transduction Program Area at Jonsson Comprehensive Cancer Center. Dr. Tamanoi is currently conducting research on nanodelivery of anticancer drugs using mesoporous silica nanoparticles. He is also exploring ways to use nanovalves to achieve controlled release of anticancer drugs. His other research topics include Signal Transduction, Protein Lipidation and Prenyltransferase Inhibitors.

Dr. Tamanoi earned his B.S. and M.S. in Biochemistry at the University of Tokyo. He received PhD in Molecular Biology at Nagoya University in 1977. He was a postdoctoral fellow at Harvard Medical School, where he worked on bacteriophage DNA replication. From 1980 to 1985, he was a senior staff investigator at Cold Spring Harbor Laboratory, where he worked on adenovirus DNA replication. From 1985 to 1993, he was an Assistant Professor and then Associate Professor at the University of Chicago, where he initiated studies on lipid modification of the Ras family proteins.

An intention of the US National Nanotechnology Initiative was to advance an interdisciplinary approach to education and science in order to promote new discoveries and technology platforms. Nowhere is it more urgent than in the evolving field of nanotoxicology. In this new field we are challenged with combining our knowledge of engineered nanoparticles, with that of molecular biology, structural biology and environmental science. Each of these areas and their associated fields of research are all "work in progress" and have their own vocabulary, protocols, and base assumptions. Finding common ground in any new cross-disciplinary area is a problem within itself. One way to bridge the disciplinary comprehension gaps is in the sharing of imagery. Current electron microscopy technology offer an extensive, and to a great extent unknown, arsenal of applications which range from site-specific imaging and analysis of bulk solid state material and biological tissues, to transmission imaging and analytical capability to the sub-atomic and molecular levels. The presentation will review 2D & 3D imaging and characterizing of nano-particle/ nanotube structures and properties; and the considerable challenge in the modeling of real nano-particle systems. Imaging and characterizing the 2D & 3D structures of tissue, cells and proteins, and the new challenge of directly visualizing the bio-nanomaterial interface. Finally we can take a glimpse into the future which promises 4D research capabilities which will enable materials and bio-chemical reactions to be studied at the highest special resolution and femto-second temporal resolution.

Mike Thompson is Business Development Manager, Nanotechnology in FEI Company; he has been actively engaged throughout his career in creating the tools to enable Nanotechnology. Following University he joined the Electron Optics division of Philips, The Netherlands as Product Manager for Transmission Electron Microscopy and subsequently lead the Philips International sales operations. He transferred to the USA as VP of the North American Philips Electron Optics sales and service division. Following the FEI / Philips merger in 1997 he moved to the West coast as VP Marketing, Sales and Service operations and helped lay the foundations of the business processes which have enabled the company's success.

In his current function he is engaged in promoting and supporting FEI's ventures and strategies to better serve the new opportunities that the Nanotechnology era presents. He has worked on the advisory boards of a number of Nanotechnology Conferences, and lectured on Nanotechnology at numerous venues. He is member of the technical advisory group which serves to advise the National Science, Engineering and Technology [NSET] committee on Nanotechnology.

Mike Thompson received his B.Sc. in Materials Science at the University of Surrey, England and his Ph.D. at Cambridge University, England.

Two critical challenges with using nanocolloids in physical processes are stability and bottom-up assembly of the particles. In contrast to larger particles, where stability and assembly are easier to bring about, in part due to a wealth of sound heuristics based on experiments and theory, for nanocolloids far fewer successful reports exist for stability and especially for assembly.

The key to dispersing and assembling nanocolloids is to control the interparticle forces. Modeling nanocolloidal forces even qualitatively is a current bottleneck for dispersion and assembly, for various reasons. New physics is required for assessing the combined roles of van der Waals (VDW) forces, electrostatic forces, and solvation-solvophobic forces. We have developed the "coupled-dipole method" (CDM) for calculating van der Waals forces for nanoparticle systems. This talk will provide a review of the traditional view of colloidal forces, give the basic physics and algorithm for the CDM, and show how we will be using CDM with other molecular simulation techniques to solve important problems for nanoscale systems.

Darrell Velegol attended West Virginia University for his BS in Chemical Engineering, and he earned his PhD in Chemical Engineering at Carnegie Mellon University in 1997 working with Professors John L. Anderson and Stephen Garoff. In 1998 Velegol won the Victor K. LaMer Award of the American Chemical Society for the best PhD in the field of Colloid & Surface Science. He continued with a post-doc in the Center for Light Microscope Imaging and Biotechnology at Carnegie Mellon, working under Professor Fred Lanni of the Biology Department. In June 1999 Velegol joined the Department of Chemical Engineering at Penn State, where he was promoted to Associate Professor in 2005. Velegol won an NSF CAREER Award in 2000, and in 2003 he led a group in winning an NSF NIRT grant on bottom-up particle assembly. Currently he works with 6 PhD students. His research investigates the fabrication of colloidal assemblies and devices, with a focus on understanding the interparticle forces and assembly process. His research group uses a wide range of experimental and modeling approaches. Velegol is a member of ACS, AIChE, AAAS, ASM, and ASEE. In calendar year 2007, Velegol is on sabbatical, with the goal of commercializing a technology from his lab.

This presentation will outline briefly the difficult balance in industry between acquiring a basic scientific understanding of materials and controlling costs and time to market.

Industrial (nano) toxicity studies usually are restricted to addressing preclinical requirements and requirements from safety regulatory authorities. These requirements are briefly outlined. Very often, companies are reluctant to invest resources and time, or simply do not have the interest, to address deviations from resolving mandated questions. The second area explored will comprise the ever-narrowing research focus on signal transduction as mechanisms of possible nanotoxicological expression. In the interest of inviting discussion, it will be proposed that careful consideration of protein and genomic array responses must be taken. Lastly, Altairnano will be introduced as an industrial company that has and continues to lead proactive studies both in-house and with federal institutions to carefully explore safety issues as well as take an in-depth look at histological tissues, beyond the scope of "what's required". Preliminary studies conducted will be presented briefly and histological highlights of that study presented. Proprietary discussion will be avoided with the understanding that sensitive material needs seclusion.

Dr. Wagner's chief interests are the structural and biochemical alterations of cellular function(s) in response to the presence of nanoparticles. Particular interests involve neurological and neuroimmunological responses to nanoparticles which may manifest in aberrant nociceptive states or the modified expression of chronic degenerative diseases. Dr. Wagner received a B.A. in Biology and Chemistry from the University of Chicago, and a doctorate in Pharmacology and Toxicology from Dartmouth Medical School. Studies (Post-Doctoral and Assistant Professor) at UCSD in Anesthesiology and Neuropathology focused on the peripheral and central etiological factors for the precipitation of chronic pain syndromes. Her efforts were the first to firmly establish the neuronal transcription of major immunological factors during tissue duress. She has also taught at all levels of education, focusing on piquing the interest of future scientists at the high school level. She recently has joined Altairnano as a Senior Research Scientist in the Life Science Business Unit.