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University of California

Center for Environmental Implications of Nanotechnology (UC CEIN)

NSF: DBI‐0830117

Annual Report Year 3

April 1, 2010 – March 31, 2011

UC Center for Environmental Implications of Nanotechnology Annual Report 2011

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TABLE OF CONTENTS

1. NSF Cover Page

2. Table of Contents 1

3. Project Summary 2

4. List of Center Participants, Advisory Boards, Participating Institutions 3

5. Quantifiable Outputs (Table 1) 9

6. Mission and Broader Impacts 10

7. Highlights 15

8. Strategic Research Plan 32

9. Research Program, Accomplishments, and Plans 35

Table 2 – NSEC Program Support 81

10. Center Diversity – Progress and Plans 82

11. Education 84

Table 3a – Education Program Participants – All 89

Table 3b – Education Program Participants – US Citizen/PR 90

12. Outreach and Knowledge Transfer 91

13. Shared and Experimental Facilities 106

14. Personnel 112

Table 4A – NSEC Personnel – All 116

Table 4B – NSEC Personnel – US Citizen/PR 118

15. Publications and Patents 120

16. Biographical Information 125

17. Honors and Awards 128

18. Fiscal Section 128

a. Statement of Unobligated Funds 128

b. Budget 128

19. Cost Sharing 153

20. Leverage 153

Table 5 – Other Support 154

Table 6 – Partnering Institutions 154

21. Current and Pending Support – PIs and Thrust Leaders


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3. Project Summary

The goal of the University of California Center for Environmental Implications of Nanotechnology (UC CEIN) is to develop a series of broad‐based predictive and integrative scientific models to understand how selected nanomaterial property‐activity relationships determine fate, transport, exposure as well as biological injury at molecular, cellular, organismal, and ecosystems levels. The integrated and multidisciplinary research effort, assisted by silico decision making tools and computational models, is assisting our understanding of how nanomaterial interactions at the nano‐bio interface and the environment could lead to hazardous outcomes and is therefore instructive of risk reduction strategies that could be undertaken to safeguard the environment. UC CEIN has successfully integrated the expertise of engineers, chemists, colloid and material scientists, ecologists, marine biologists, cell biologists, bacteriologists, toxicologists, computer scientists, biostatisticians, and social scientists into a predictive scientific platform that inform us about possible ENM hazards and how through exposure reduction, lifecycle analysis and safe‐by‐design strategies to reduce the environmental impact of nanotechnology.

The work of the Center is carried out by seven interdisciplinary research groups (IRGs): IRG 1: Acquisition of ENM compositional and Combinatorial Libraries and Physical‐chemical Characterization; IRG 2: Studying ENM Interactions at the Molecular, Cellular, Organ, and System Levels; IRG 3: Organismal and Community Ecotoxicology; IRG 4: Nanoparticle Fate and Transport; IRG 5: High‐Throughput Screening (HTS), Data Mining, and Hazard ranking to assist in vivo studies; IRG 6: Modeling of ENM distribution in the environment, Multimedia modeling, Development of in silico Decision‐making Tools for hazard ranking and establishing Quantitative‐Structure Relationships; IRG 7: Risk Perception of Potential Environmental Impacts of Nanotechnology.

Now in the third year of operation, the UC CEIN has impacted national and international understanding and decision‐making in the areas of NanoEHS research, protocol development, knowledge dissemination, and contributions to the regulatory agencies. Currently, the Center engages in 38 research projects that are supported by three major support cores and a Center administration. One of the key scientific accomplishments of the Center to date has been the synthesis, characterization, and implementation of a metal oxide (TiO2, CeO2, and ZnO) nanoparticle library across all Center projects, leading to multidisciplinary and integrated efforts that have resulted in the generation of a major body of knowledge on how these materials impact a wide range of environmental lifeforms. Further expansion of the nanomaterial libraries have now yielded over a 100 compositional or combinatorial variations to study through the performance of cellular high throughput screening and the implementation of high content screening in zebrafish embryos. The screening efforts that evaluate sublethal and lethal injury outcomes is being used for prioritizing the study of a range of organisms in freshwater, marine, and terrestrial environments, including their fate and transport that leads to exposure under these conditions. High content data generation assisted by heatmaps, self organizing maps and in silico feature selection methodology has allowed hazard ranking and a nano‐SAR classification to be developed for identifying hazardous material properties. Data collection and analysis on environmental risk perception and multimedia modeling of air, water, and soil exposure conditions are ongoing. The Education and Outreach programs disseminate the knowledge generation to our students, the public and national agencies; UC CEIN has had a broad impact on the scientific, educational, and policy communities nationally and internationally.

In the coming year, we will continue our predictive scientific investigation and modeling of a progressively wider range of ENMs and their impact on the environment. We are co‐sponsoring ICEIN 2011, are assisting in EU/US collaboration and we will continue to play a leading role in national and international Nano EHS forums. We will also expand our collaboration and interaction with State and Federal agencies.


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4. Center Participants, Advisory Boards, and Participating Institutions Center Participants Participants Receiving Center Support Faculty: Kenneth Bradley UCLA Assistant Professor, Microbiology Jeffrey Brinker University of New Mexico/Sandia Professor, Chemical/Nuclear Engineering Bradley Cardinale UC Santa Barbara Assistant Professor, Ecology Evolution, Marine Biology Gary Cherr UC Davis Professor, Environmental Toxicology/Nutrition Yoram Cohen UCLA Professor, Chemical Engineering Curtis Eckhert UCLA Professor, Environmental Health Sciences William Freudenberg UC Santa Barbara Professor, Environmental Studies and Sociology Jorge Gardea‐Torresdey University of Texas, El Paso Professor, Chemistry Hilary Godwin UCLA Professor, Environmental Health Sciences Robert Haddon UC Riverside Professor, Chemistry Barbara Herr Harthorn UC Santa Barbara Associate Professor, Women’s Studies/Anthropology Mark Hersam Northwestern University Professor, Materials Science & Engineering Eric Hoek UCLA Associate Professor, Civil & Environmental Engineering Patricia Holden UC Santa Barbara Professor, Environmental Microbiology Milind Kandlikar University of British Colombia Assistant Professor, Institute for Global Issues Arturo Keller UC Santa Barbara Professor, Environmental Biogeochemistry Hunter Lenihan UC Santa Barbara Associate Professor, Marine Biology Alex Levine UCLA Assistant Professor, Chemistry and BioChemistry Shuo Lin UCLA Professor, Molecular, Cell, & Developmental Biology Lutz Madler University of Bremen Professor, Materials Science Timothy Malloy UCLA Professor, Law Edward McCauley UC Santa Barbara Professor, Ecology, Evolution, Marine Biology Andre Nel UCLA Professor, Medicine; Chief, Division of NanoMedicine Roger Nisbet UC Santa Barbara Professor, Ecology, Evolution, Marine Biology Robert Rallo Universitat Roriv i Virgili/UCLA Associate Professor, Chemical Engineering Theresa Satterfield University of British Colombia Associate Professor, Institute of Resources Joshua Schimel UC Santa Barbara Professor, Ecology, Evolution, Marine Biology Ponisseril Somasundaran Columbia University Professor, Materials Science Galen Stucky UC Santa Barbara Professor, Chemistry and Biochemistry Donatello Telesca UCLA Assistant Professor, Biostatistics Sharon Walker UC Riverside Assistant Professor, Chemical and Environmental Eng. Jeffrey Zink UCLA Professor, Chemistry and Biochemistry Research Staff: Raven Bier UC Santa Barbara Irina Chernyshova Columbia University Robert Damoiseaux UCLA Anna Davison UC Santa Barbara Helen Dickson UC Santa Barbara Aoergele Fnu UCLA Bryan France UCLA Jennifer Gowan UC Santa Barbara Taimur Hassan UCLA Sean Hecht UCLA Susan Jackson UC Davis Zhaoxia Ivy Ji UCLA Xingmao Jiang Sandia National Labs Ya‐Hsuan Liou UC Santa Barbara


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Marianne Maggini UC Santa Barbara Huan Meng UCLA Robert Miller UC Santa Barbara Delia Milliron Lawrence Berkeley National Laboratory Taleb Mokari Lawrence Berkeley National Laboratory Erik Muller UC Sana Barbara Jose Peralta‐Videa University of Texas, El Paso Dad Roux‐Michollet UC Santa Barbara David Schoenfeld UCLA Carol Vines UC Davis Hongtuo Wang UC Santa Barbara William Wooten UCLA Tian Xia UCLA Postdoctoral Researchers: Mafalda Baptista UC Santa Barbara Rafaella Buonsanti UCLA/Lawrence Berkeley National Laboratory Bryan Cole UC Davis Gwen D’Arcangelis UC Santa Barbara Guadalupe De La Rosa University of Texas, El Paso Elise Fairbairn UC Davis Xiaohua Fang Columbia University Yuan Ge UC Santa Barbara Saji George UCLA Nalinkanth Ghone UCLA Debraj Ghosh UCLA Yongsuk Hong UC Santa Barbara Chia‐Hung Hou UC Santa Barbara Angela Ivask UCLA Xue Jin UCLA Mikael Johansson UC Santa Barbara Sanaz Kabehie UCLA Irina Kalinina UC Riverside Myungman Kim UCLA Hiroaki Kiyoto UC Santa Barbara Tin Klanjscek UC Santa Barbara Chris Knoll UC Santa Barbara Konrad Kulacki UC Santa Barbara Minghua Li UCLA Sijie Lin UCLA Rong Liu UCLA Martha Lopez University of Texas, El Paso Cecile Low‐Kam UCLA Milka Montes UC Santa Barbara Sumitra Nair UCLA Sandip Niyogi UC Riverside Manuel Orosco UCLA Anton Pitts University of British Columbia Suman Pokhrel University of Bremen John Priester UC Santa Barbara Aditi Singhal UC Santa Barbara Elizabeth Suarez UCLA Won Suh UC Santa Barbara


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Reginald Thio UC Santa Barbara Raja Vukanti UC Santa Barbara Xiang Wang UCLA Haiyuan Zhang UCLA Lijuan Zhao University of Texas, El Paso Yan Zhao UCLA Graduate Students: Khadeeja Abdullah UCLA Adeyemi Adeleye UC Santa Barbara Hayley Anderson UCLA Barbora Bakajova UC Santa Barbara Lynn Baumgartner UC Santa Barbara Christian Beaudrie University of British Columbia Samuel Bennet UC Santa Barbara David Boren UCLA Olivier Brun UC Santa Barbara Benjamin Carr UC Santa Barbara Savanna Carson UCLA Eunshil Choi UCLA Kabir Chopra UCLA Indranil Chowdhury UC Riverside Kristin Clark UC Santa Barbara Mary Collins UC Santa Barbara Alyssa de la Rosa University of Texas, El Paso Laura De Vries University of British Columbia Cassandra Engeman UC Santa Barbara Daniel Ferris UCLA Allison Fish UC Santa Barbara Thomas Glaspy UCLA Shannon Hanna UC Santa Barbara Jose Hernandez‐Viezcas University of El Paso, Texas Ryan Honda UC Riverside Allison Horst UC Santa Barbara Carlin Hsueh UCLA Daniel Huang UC Santa Barbara Kathryn Leonard UCLA Zongxi Li UCLA Monty Liong UCLA Haoyang Haven Liu UCLA Catalina Marambio‐Jones UCLA David McGrath UCLA John Meyerhofer UC Santa Barbara Randy Mielke UC Santa Barbara Sudhir Paladugu UC Santa Barbara Trina Patel UCLA Satish Ponnurangam Columbia University April Sawvell UC Santa Barbara Alia Servin University of Texas, El Paso Sharona Sokolow UCLA Louise Stevenson UC Santa Barbara Sirikarn Surawanvijit UCLA Derrick Tarn UCLA


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Courtney Thomas UCLA Michael Tsang UCLA Jessica Twining UC Santa Barbara Pria Vytla UC Santa Barbara Rebecca Werlin UC Santa Barbara Tristan Winneker UC Santa Barbara Kimberly Worsley UC Riverside Sijing Xiong Nanyang Technological University Min Xue UCLA Kristin Yamada UCLA Yichi Zhang UC Santa Barbara Dongxu Zhou UC Santa Barbara Undergraduate Students: Nicolai Archuleta UC Santa Barbara Rebecca Britt Armenta University of Texas, El Paso Bernice Chan UCLA Gwen Christiansen UC Santa Barbara Maia Colyar UC Santa Barbara Jon Conway UC Santa Barbara Stephen Crawford UC Santa Barbara Israel Del Toro University of Texas, El Paso Vivian Do UCLA Janel Feige UCLA Ryo Furukawa UCLA Arjan Gower UC Santa Barbara Edward Hu UC Santa Barbara James Kim UC Riverside Soomin Kim UC Santa Barbara Casey Leavitt UC Santa Barbara Leuh Yang Liao UCLA Erica Linard UC Santa Barbara Angela Liu UCLA Kristin Matulich UC Santa Barbara Ariel Miller UC Santa Barbara Alex Moreland UC Santa Barbara Fabiola Moreno University of Texas, El Paso Kaysha Nelson UC Santa Barbara Michelle Oishi UCLA Leanne Paragas UCLA Calvin Parshad UCLA Scott Pease UC Santa Barbara David Pereira UC Santa Barbara Gabriel Rubio UC Santa Barbara Esther Shin UC Davis Tiffany Takade UC Santa Barbara Nancy Tseng UC Santa Barbara Kari Varin UCLA Celia Veg‐Herrera UCLA Christina Wong UCLA High School Students (Interns): Courtney Kwan UC Santa Barbara


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Staff/Administration: David Avery UCLA John Chae UCLA Mariae Choi UCLA Julie Dillemuth UC Santa Barbara Kristin Duckett UC Santa Barbara Vi Tuong Huynh UCLA Catherine Nameth UCLA Nancy Neymark UCLA Affiliated Participants, Not Receiving Center Support Faculty: Carolyn Bertozzi UC Berkeley/Lawrence Berkeley Lab Professor, Chemistry, Molecular/Cell Biology Freddy Boey Nanyang Technological University Professor, Materials Science Engineering Kenneth Dawson University College Dublin Professor, Physical Chemistry Francesc Giralt Universitat Rovira I Virgili Professor, Chemical Engineering Jordi Grifoll Universitat Rovira I Virgili Associate Professor, Chemical Engineering Joachim Loo Nanyang Technological University Assistant Professor, Materials Engineering Nick Pidgeon Cardiff University Professor, Applied Psychology Graduate Students: Xinxin Zhao Nanyang Technological University External Science Advisory Committee Pedro Alvarez Rice University Professor, Engineering Ahmed Busnaina Northeastern University Professor, Engineering; Director, HRNM Sharon Dunwoody University of Wisconsin‐Madison Professor, Journalism/Mass Communication Menachem Elimelech Yale University Professor, Chemical Engineering C. Michael Garner Intel Corporation Program Manager, Emerging Materials Res. James Hutchison University of Oregon Professor, Assoc. VP, Research Fred Klaessig Pennsylvania Bio Nano Systems Julia Moore Woodrow Wilson International Center Deputy Director, PEN Kent Pinkerton UC Davis Director, Center for Health/Environment David Rejeski Woodrow Wilson International Center Director, PEN Ron Turco Purdue University Professor, Agronomy Isiah Warner Louisiana State University Professor, Environmental Chemistry Jeff Wong Department of Toxic Substances Control Deputy Director, Science Academic Participating Institutions Cardiff University Centro de Investigacion y de Estudios Avanzados del Instituto Politechnico Nacional (CINVESTAV) Columbia University Instituto Nacional de Salud Publica (INSP) Nanyang Technological University Universitat Rovira I Virgili University of Bremen University of British Colombia University of California, Los Angeles University of California, Santa Barbara University of California, Davis University of California, Riverside University College Dublin


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University of New Mexico University of Texas, El Paso Non Academic Participating Institutions California Science Center Lawrence Berkeley National Laboratory Lawrence Livermore National Laboratory Sandia National Laboratory Santa Monica Public Library

Table 1: Quantifiable Outputs - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Outputs Reporting Year -4 Reporting Year -3 Reporting Year -2 Reporting Year -1 Reporting Year Total

Publications Resulted From NSEC Support

In Peer Reviewed Technical Journals 0 0 12 27 48 87

In Peer Reviewed Conference Proceedings 0 0 0 1 2 3

In Trade Journals 0 0 0 0 2 2

With Multiple Authors 0 0 12 27 50 89

Multiple Authors: Co-Authored With NSEC Faculty 0 0 12 27 50 89

NSEC Technology Transfer

Inventions Disclosed 0 0 0 0 0 0

Patents Filed 0 0 0 0 0 0

Patents Awarded 0 0 0 0 0 0

Patents Licensed 0 0 0 0 0 0

Software Licensed 0 0 0 0 0 0

Spin-off Companies Started (if applicable) 0 0 0 0 0 0

Degrees to NSEC Students

Bachelor's Degrees Granted 0 0 0 0 3 3

Master's Degrees Granted 0 0 0 2 1 3

Doctoral Degrees Granted 0 0 0 1 2 3

NSEC Graduates Hired by

Industry 0 0 0 0 1 1

NSEC participating firms 0 0 0 0 0 0

Other U.S. Firms 0 0 0 0 1 1

Government 0 0 0 0 1 1

Academic Institutions 0 0 0 2 0 2

Other 0 0 0 0 0 0

Unknown 0 0 0 0 0 0

NSEC Influence on Curriculum (if applicable)

New Courses Based on NSEC Research 0 0 0 1 2 3

Courses Modified to Include NSEC Research 0 0 0 6 3 9

New Textbooks Based on NSEC Research 0 0 0 0 0 0

Free-standing Course Modules or Instructional CDs 0 0 0 0 18 18

New Full Degree Programs 0 0 0 0 0 0

New Degree Minors or Minor Emphases 0 0 0 0 0 0

New Certificate 0 0 0 0 0 0

Information Dissemination/Educational Outreach

Workshops, Short Courses to Industry 0 0 0 1 1 2

Workshops, Short Courses to Others 0 0 0 2 2 4

Seminars, Colloquia, etc. 0 0 49 211 185 445

World Wide Web courses 0 0 3 1 2 6

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6. Mission Statement The mission of the University of California Center for Environmental Implications of Nanotechnology (UC CEIN) is to ensure that nanotechnology is introduced in a responsible and environmentally compatible manner, thereby allowing the US and International Communities to leverage the benefits of nanotechnology for global economic and social benefit. This mission is being accomplished by developing a series of decision tools based on models of predictive toxicology and risk ranking premised on selected nanomaterial properties that determine fate, transport, exposure, and biological injury mechanisms at cellular, tissue, organism, and population levels. Since its founding in September 2008, the UC CEIN has successfully integrated the expertise of engineers, chemists, colloid and material scientists, ecologists, marine biologists, cell biologists, bacteriologists, toxicologists, computer scientists, biostatisticians, and social scientists into a predictive scientific platform that informs us about possible ENM hazards and how through exposure reduction, lifecycle analysis and safe‐by‐design strategies we can reduce the environmental impact of nanotechnology

The key components of the predictive and inter‐related scientific models include: (i) the establishment of nanomaterial libraries based on a consideration of production volumes and the material types most likely to come into contact with the environment; (ii) nanomaterial distribution to the environment as determined by modes of release, physicochemical and transport properties, contact and interaction with biological receptors, and bioaccumulation; (iii) representative ecological life forms serving as early sentinels to monitor the spread and bio‐accumulation of hazardous nanomaterials; (iv) mesocosm, population, and dynamic energy budget theory of freshwater, marine, and terrestrial environments; (v) biological and high throughput screening assays using cells, bacteria, embryos, etc., to generate information about hazardous ENM properties (included in compositional and combinatorial ENM libraries) that can help to execute and plan environmental health effect studies on increasing complex life forms in the environment; and (vi) a series of in silico decision making tools and computational models to predict nanomaterial interactions at the nano‐bio interface and in the environment. These research activities are being combined with educational programs to inform the public, future generations of scientists, public agencies, and industrial stakeholders of the importance of safe implementation of nanotechnology in the environment. The overall impact will be to reduce uncertainty about the possible consequences of nanomaterials in the environment, while at the same time providing guidelines for their safe design and use to prevent environmental hazard. Broader Impacts Traditional and current toxicity testing in humans and the natural environment is heavily dependent on a complex set of whole‐animal‐based toxicity testing strategies. This approach, while a time‐honored hazard assessment tool, is unlikely to handle the rapid pace at which nanotechnology‐based enterprises are generating new materials. The UC CEIN is addressing these challenges of scale by using a scientific platform that can perform high content and high throughput screening to generate the knowledge domains that are required to make predictions about the impact of nanotechnology on the environment. This knowledge is also being used for making predictions and implementing the safe design of nanomaterials. The UC CEIN’s creation of in silico decision making tools is assisting hazard ranking and risk predictions of nanomaterial impacts on the environment for the global society. Our educational and outreach activities are being used as powerful portals for the dissemination of our research findings and predictions to the scientific and industrial communities. Our outreach activities are informing both experts and the public at large about the safety issues surrounding nanotechnology, including hazard assessment, risk perception, risk reduction strategies and safe‐by design production methods.


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Significant Advances since April 1, 2010 The significant advances during the third year were to consolidate the startup efforts and implementation of research methodology, protocols, approaches and projects launched during launched during the center's first 21 months into an impactful, multidisciplinary and educational work product. One major accomplishment was the synthesis, characterization and implementation of a metal oxide (TiO2, CeO2 and ZnO) nanoparticle library across all Center projects, leading to a series of multidisciplinary and integrated efforts that has resulted in the generation of a major integrated body of knowledge regarding the behavior of these materials in the environment. The major findings of this harmonization effort, as reviewed in our ACS Nano focus review (volume 5, p 13‐20, 2011) by multiple authors across five IRGs, are that it is possible to develop reproducible physicochemical characterization and assessment of the state of dispersion of metal oxide nanoparticles in multiple biological and environmental use media, including tissue culture media, bacterial broth, and eight representative environmental media. Not only were we able to determine in our fate and transport studies that the state of aggregation and dispersal of these particles determine the exposure of pelagic versus benthic organisms, but were also able to develop a number of dispersing agents that could be used for conducting biological experiments to compare the effects of agglomerated and dispersed nanoparticles. In addition, we have successfully implemented high throughput dynamic light scattering to assist us in determining the kinetics of agglomeration and stability of the colloidal suspensions in the various media with considerable ease. We have demonstrated that through the implementation of high throughput screening that makes use of a robotic system and assessment of appropriate injury pathways in mammalian tissue culture cells and bacteria that it is possible to perform hazard ranking of metal oxide nanoparticles. Moreover, we were able to compare the in vitro screening results with the biological outcome in a variety of environmental lifeforms, including phytoplankton, oyster embryos, algae and zebrafish embryos (see details below). On average, ZnO nanoparticles tended to be more toxic than TiO2 and CeO2 by a mechanism that involves extensive particle dissolution. Moreover, we demonstrated that through iron doping of the ZnO nanoparticles that it is possible to change the particle matrix to yield a library of slower dissolving particles that are toxic in tissue culture cells, bacteria and zebrafish embryos. This constitutes an important example of a safe‐b‐ design feature that were also tested in other environmental studies. While TiO2 was relatively devoid of toxicity under dark and visible light conditions, these nanoparticles did show phytoplankton toxicity under UV exposure conditions, indicating that photo oxidation could be an important consideration under bright sunlight conditions. This sparked the deliberate synthesis and characterization of an Fe‐doped TiO2 nanoparticle library that can be studied under less toxic year‐visible light conditions. Because of the high‐volume data sets that are being generated during high throughput screening, we were also able to develop in silico software programs that have allowed us to generate heat maps, self‐organizing maps and some of the first QSARs for linking nanomaterial properties to adverse biological outcome. In addition to this major integrative activity leading to a series of work products across the center, work undertaken during year three have yielded the following major accomplishments that are cited in bullet point format:

Testing of ZnO and TiO2 across ecosystem types has shown: (i) that ZnO under most conditions is toxic to most sentinel species, mostly due to the Zn2+ ion exposure rather than the ENMs themselves; (ii) that TiO2 is rarely toxic unless there is photoactivation, under which circumstances it could become and may have an impact on phytoplankton; (ii) that ceria is rarely


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toxic but may generate genotoxicity in plants; (iv) the presence of organic matter in marine sediments greatly decreases the bioavailability and toxicity of MeOs in marine ecosystems; (v) that uptake and biotransformation by plants and bio‐filtration by marine suspension feeders substantially modifies or sequesters MeO ENMs making them less bioavailable in some of organisms but could lead to bioaccumulation in some grazers and predators; (vi) that community biodiversity can influence the bioavailability of ENMs in food webs contained within mesocosms (IRG 3)

In studies addressing the mobility, persistence, bioavailability and reactivity of NPs in actual environmental media, IRG4 has found that : (i) ZnO is likely to dissolve rapidly (within days) in most natural conditions, so that the risk is mostly due to the Zn2+ rather than the NPs themselves; (ii) the MeOs aggregate rapidly in seawater and other high ionic strength media, but are much more stable in freshwater and other media high in NOM and low ionic strength; (iii) the presence of NOM plays a major role in preventing attachment of the MeO NPs to mineral surfaces; (iv) filtration of the NPs in groundwater and sediments may occur if they aggregate significantly, but mostly due to straining as opposed to attachment to the porous media; (v) the mobility of the MeO NPs is greatly increased by the presence of NOM, whether in seawater, freshwater or groundwater; (vi) the photoactivity of the MeO NPs is a strong function of surface chemistry, followed by the effect of the surrounding media; and (vii) NP morphology plays an important role in controlling aggregation, with results so far indicating that a flat, plate‐like morphology aggregates much faster than a spherical particle (IRG 4).

The CEIN nanomaterial libraries were expanded in 2010‐2011 to now include more than 100 different particle types (IRG1). These include a range of titania, ceria, and silica sizes and shapes (cubes, rods and spheres), a silver nanoparticle library that includes monodispersed 10nm and polydispersed 57 nm particles; single walled carbon nanotubes (commercial and purified), platinum, and a silica nanoparticle library that includes amorphous, mesoporous, fumed, and crystalline silica polymorphs (IRG 1).

IRG 1 has expanded its characterization capabilities to include surface area and pore volume measurements by using gas absorption/desorption isotherms and BET analysis. A high throughput instrument that allows 4, concurrent measurements is currently being used by the CEIN to study surface area and pore volumes of library materials (IRG 1).

Implementation of high content zebrafish embryo screening to supplement the in vitro (cellular) HTS, with the capability to compare the multi‐parametric cellular responses to Ag, Au, Pt, Al2O3, SiO2, ZnO and Q‐dot (CdSe/ZnS) nanoparticles to the survival, hatching, and morphological defects that developing zebrafish embryos. Our data have shown that there is good agreement between the in vitro hazard ranking and zebra embryotoxicity, with the exception of nano‐Ag that proved more deleterious to the embryos than to mammalian tissue culture cells. We have now implemented a zebra fish gill cell line to determine whether there are differences in the mechanisms of cellular toxicity using mammalian and fish cells (IRG5).

Completed all measurement interpretations related to simple trophic transfer study (bacteria to protozoans) of Cd(II) versus CdSe QDs where significant bioaccumulation in bacteria and biomagnification to protozoa were observed. Used combined STEM and growth modeling to reveal digestion impedance visualized in STEM images, and defined major discovery of this experiment as one of differential digestion due to feeding on QD‐laden bacteria. The published manuscript is one of the first comprehensive papers showing biomagnification of engineered nanomaterials (IRG2).

Developed dynamic energy budget model of effects of Cd(II) on bacterial growth, using growth curve data collected experimentally for CdSe QD and Cd(II) growth inhibition studies published


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in 2009. Showed that assumptions involving reactive oxygen species (ROS) affecting controls improve the model fit to data. Completed calibration of model for new data generated of optical density versus cell counts, using these results to calibrate data at high cell density regions of growth curves. Improved model using new assessments of lag times (IRG2).

In the evaluation of sea urchin embryo toxicity in response to the metal oxides (CeO2, TiO2, ZnO), ZnO was toxic to developing embryos, causing developmental abnormalities at very low levels with dose‐ and exposure time‐dependencies, but without an effect on the developmental stages. Curiously, Fe‐doped ZnO showed comparable toxicity in sea urchin embryos as the non‐doped ZnO particles, indicating that the accelerated particle agglomeration in this environmental medium does not change the dissolution characteristics to similar extent as in Holtfreter's medium or mammalian tissue culture media (IRG 2).

Selected and recruited assays for evaluating sublethal effects of ENMs on bacteria for translation to HTS, showing the tremendous potential for HTS of bacteria for assessing potential environmental impacts of ecosystem relevance (IRG 2 and IRG5).

Based on the newly developed in silico feature selection methodology, a nano‐SAR classification was developed for cytotoxicity of metal and metal oxide nanoparticles. This work revealed that atomization energy, the period of the metal, the nanoparticle volume (in solution), and the primary particle size were fundamental descriptors that enabled correlation to cytotoxicity with a high level of accuracy. The classification‐based nano‐SAR enables one to identify decision boundaries that are crucial for use in hazard ranking of nanoparticles (IRG 6). We are expanding on this concept by introducing a wide range of metal oxide nanoparticles in which we will include above features as well as information about the band gap distribution to study the relationship to HTS parameters for cytotoxicity in mammalian cells and bacteria (IRG5 and IRG6).

Lung epithelial cells were transduced with the luciferase reporter genes that reflect the transcriptional activation of a number of intracellular signaling cascades. The functionality of the stable transformed cell lines was verified with positive stimuli. These cell lines have been scaled up to HTS format for screening with eight standard reference materials (gold, silver, platinum, aluminum, zinc, silica, iron, and CdSe/ZnS Quantum Dots). The analysis of the signaling‐pathway response signatures identified two distinct groups of meta‐clusters corresponding to (i) sub‐lethal pro‐inflammatory responses that are possibly related to ROS generation, and (ii) lethal genotoxic responses due to exposure to ZnO and Pt nanoparticles at high concentrations. The most significant meta‐cluster was related to DNA damage and included the cell signaling responses to ZnO and Pt nanoparticles. The results of the study suggest that SOM analysis enables expanded knowledge extraction from HTS nanoparticle data with clear visualization of patterns and clustering of cell responses. The SOM approach was utilized by IRG six to develop predictive quantitative‐structure relations (IRG 5).

Preliminary results using complex network theory methods to study the relationships between signaling pathways and observed toxicity identified the presence of three well‐differentiated communities (clusters). The use of association rules has also been adapted to identify significant relationships among the signaling pathways and the cytotoxicity. This analysis based on both RAW and BEAS‐2B mammalian cell lines, for example, indicates the presence of a hierarchical activity‐activity pattern where sub‐lethal effects such as ROS generation and the intracellular Ca2+ flux are strongly related to lethal effects (e.g. cell membrane damage and cell death) (IRG 6).

Data collection on environmental risk perception was completed in a two‐phase design that looked at the public perceptions of air, water, and soil (phase 1) and their interaction with ENMs (phase 2). The study was designed with input from IRGs 1‐4 to determine which ENMs to focus


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on, to ensure the validity of distinctions, and to ensure instrument conformity to ecologists' views of environmental media. Data are currently being analyzed in preparation for publication (IRG 7).

Impacts of Education and Outreach Programs A major goal of the UC CEIN is to train the next generation of nano‐scale scientists, engineers, and regulators to anticipate and mitigate potential future environmental hazards associated with nanotechnology. Our educational programs are developed to broaden the knowledge base of the environmental implications of nanotechnology through academic coursework, world‐class research, training courses for industrial practitioners, public outreach, and a journalist–scientist communication program.

Significant progress has been made on several Education and Outreach goals that enhance our ability to have a broader impact on the scientific, educational, and policy communities both nationally and internationally. These include: (i) activities that integrate across IRGs to ensure a unified and effective approach to mentoring and training both students and postdoctoral researchers; and (ii) activities focusing on communicating the findings of the UC CEIN to a broader audience and varied constituencies with the goal to framing these findings to provide a scientific basis for related policy and regulatory decisions. Key accomplishments in the past year are detailed in section 11, highlights of which include:

UC CEIN taking a national leadership role in Nano‐EHS through participation in the Nano2 initiative exploring the NNI's vision for Nanotechnology in the next 10 years. Additionally, PI Nel is a member of the Nano EHS Russia‐US Bilaterial Presidential Commission of Experts and several Center faculty have represented our Center at high profile conferences over the past year.

Offered three leadership workshops for Center students and postdocs on the topics of: High Throughput Screening and Analysis of Large Data Sets (May 2010); Communicating your Science to the Public (Feb 2011); and the Academic Job Search (March 2011).

Made publically available the first online graduate level short course on Dynamic Energy Budget Theory via iTunes U.

Adapted our exisiting Nanotoxicology Capstone course to an online Nanoecotoxicology course that is currently being pilot tested by graduate level students in Mexico. The course will be made available to all Center members, and eventually will be offered to any of our educational partners.

Co‐hosted Nanotechnology VI: Progress in Protection (October 13, 2011 with California Department of Toxic Substances Control) ‐ a day‐long workshop on the Environmental Health and Safety of Engineerined Nanomaterials.

Hosted and co‐sponsored the 2010 International Conference on the Environmental Implications of Nanotechnology (with CEINT) at UCLA in May 2010. Planning underway for ICEIN 2011 to be held at Duke University in May 2011.

Formalized partnerships with the California Science Center and the Santa Monica Public Library to organize a series of public outreach events. Contributed additional public outreach materials to the Santa Barbara Public Library, the Brentwood School, and the UCLA California NanoSystems Institute.

Partnered with the California Department of Toxic Substances Control to provide an evaluation of the recent mandatory statewide call‐in for carbon nanotubes. Created a California Nano EH&S


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Working group which has developed and is testing guidance for the safe handling of ENMs in academic settings.

In Summer 2010, a web survey of 424 non S&E, nanotoxicology, and regulator experts was completed to asses views of ENM risks and regulations. The results of this study will provide a vital comparative framework for UC CEIN public and industry risk perception studies. The study supports the need for the development of decision‐analytic tools (risk‐ranking, multi‐criteria decision analysis, and control banding) adapted to decision making re: environmental risks of ENMs.

7. Highlights Research, Education, and Outreach Highlights follow.

Nanomaterials in the Environment: From Materials to High-Throughput Screening to Organisms

Courtney R. Thomas, Saji George, Allison M. Horst, Zhaoxia Ji, Robert J. Miller, Jose R. Peralta-Videa, Tian Xia, Suman Pokhrel, Lutz Madler, Jorge L. Gardea Torresdey, Patricia A. Holden, Arturo A. Keller, Hunter S. Leniahn,

Andre E. Nel, and Jeffrey I. Zink. UC Center for Environmental Implications of Nanotechnology

Libraries of materials are first characterized to establish theproperties of the materials. After HTS toxicity screening, thedata can be analyzed and displayed as heat and self-organizing maps, with fluorescence signals seen in yellowand red corresponding to increased toxicity. At the sametime, the mechanism of toxicity will be determined and linkedto the physicochemical properties of nanomaterials.Nanomaterials are then prioritized with regard to furthertoxicity screening. Model organisms in various trophic levelswill be used to examine the toxicity of nanomaterials. This -information will be used to build the structure-activityrelationships established using cell studies and confirmed invivo; new materials can be synthesized based on safedesign principles. These new materials are added to thecombinatorial libraries, and tested to verify the hypothesizedreduced toxicity.

NSF: DBI-0830117

ACS Nano 5(1) p.13-20, 2011

The reduction in the band gap energy of TiO2 with increasing Fe loading had no effect in the crystallinity and the homogeneousdistribution of the nanoparticles. Photochemical studies showed that band gap was reciprocally tuned proportional to the Fe contentand the photo-oxidation increased with Fe loading during irradiation. The cytotoxic and ROS production in the macrophage cell lineshowed increased oxidant injury and cell death with a decreased band gap energy. These findings reflect the potential of Fe-TiO2 togenerate adverse effects in humans and the environment during high intensity light exposure. (Work submitted for publication).

Effects of near-visible light irradiation to the Fe doped TiO2 exposed cellsS. George ς,δ, S. Pokhrel¥, Z. Jiχ, B. Hendersonχ, Tian Xia ς, δ, L. Li ς, δ, J. I. Zinkχ, A. E. Nel ς, δ, Lutz Mädler¥,δ

ςDepartment of Medicine-Division of NanoMedicine, University of California, Los Angeles, California, USA, δCalifornia NanoSystems Institute at University of California, Los Angeles, California, USA, ¥Foundation Institute of Materials Science (IWT), Department of Production Engineering, University of Bremen,

Germany, χDepartment of Chemistry and Biochemistry, University of California, Los Angeles, California, USA

IRG 1, L. Mädler, S. Pokhrel, J. I. Zink, Z. Ji, B. Henderson

IRG 2, A. E. Nel, S. George, T. Xia, L. Li

NSF: DBI-0830117

Effects of Zn-containing compounds on sea urchin development. Elise A. Fairbairn, Arturo A. Keller, Lutz Mädler, Dongxu Zhou, Suman Pokhrel, Gary N. Cherra

UC Center for Environmental Implications of Nanotechnology

Newly fertilized embryos were exposed to Zn-containing compounds (ZnO nanomaterial, or 10% Fe-doped ZnO NM) until Control embryos reached the pluteus stage (approximately 96 hours after fertilization), then fixed and assessed for normal/ abnormal development. Fe-doped ZnO NM was less soluble than pure ZnO NM in seawater. At concentrations near those used in the toxicity assays, Fe-doped ZnO NM reached ~80% dissolution, compared to nearly 100% dissolution with the pure ZnO NM. However, in contrast to the reduced toxicity observed in the in vitro cell culture system, we observed no significant difference between toxicity with Fe-doped ZnO NM and the pure ZnO NM in our sea urchin developmental bioassay.

NSF: DBI-0830117

J. Hazardous Materials (2011). Accepted.

Biomagnification of CdSe QDs: Pseudomonas to TetrahymenaR. Werlin, J. H. Priester,R. E. Mielke, S. Krämer, S. Jackson, P. K. Stoimenov, G. D. Stucky, G. N. Cherr, E. Orias, P. A. Holden

UC Center for Environmental Implications of Nanotechnology

Biomagnification of pollutants is a pinnacle concern in ecotoxicology. Here, using basal organisms in most food webs, the potential for biomagnifcation of an engineered nanomaterial was shown anew. A common protozoan predator, Tetrahymena thermophila, when feeding exclusively on its bacterial prey that had internalized quantum dots (QDs), Pseudomonas aeruginosa, biomagnified cadmium in the form of intact QDs (bar graph, with dark bars for QD cadmium). Intact QDs distributed throughout the predators, but ultimately stunted digestion, leaving undigested early food vacuoles (eFV) packed with QD-laced bacteria (right, arrow and triangle). These protozoans, owing to swimming cessation, are particularly susceptible to predation which could increase trophic transfer.

NSF: DBI-0830117

(Werlin, et al. 2011, Nat. Nano. 6, 65-71)

NM TiO2 reduces marine phytoplankton population growth rates in high UV light levels because ROS is generated. ROS causes multiple forms of cytotoxicity.

We are investigating the effects of TiO2 on population growth rates of marine phytoplankton, and DEB modeling was used to quantify effect parameters, including no-effect concentrations for four common species of coastal phytoplankton.

In experiments exposing phytoplankton to TiO2 NPs with (red dots) and without (black dots). environmentally relevant UV light exposure, TiO2 was shown to be toxic only with UV exposure. The toxicity levels were at relatively high concentrations (>4ppm).

Population tests will proceed with the next generation of CEIN NP libraries.

NSF: DBI-0830117

Robert J. Miller1, Hunter S. Lenihan2, Scott Pease2, Edward Hu21Marine Science Institute, UCSB 2Bren School of Environmental Science and Management, UCSB

Submitted to Environmental Science & Technology.

Shannon Hanna1, Robert J. Miller2, Hunter S. Lenihan1, Erik Muller3, Roger Nisbet31Bren School of Environmental Science and Management, UCSB 2Marine Science Institute, UCSB 3Department of Ecology, Evolution

and Marine Biology, UCSB

Marine mussels are used extensively in marine pollution monitoring programs (e.g., The CA Mussel Watch program) to test for the presence and bioavailability of contaminants in marine systems. Mussels are a sentinel species for another ecosystem service, biofiltration by suspension feeders. Mussels are also critical links between phytoplankton and benthic consumers in coastal reef food webs.

1000+ mussels were exposed to ZnO ENMs over a 3 month period. Early data analysis shows exposure to ZnO ENMs inhibit mussel growth with increasing concentrations, but may promote growth at low levels. Additionally, mussels show bioaccumulation of Zn in soft tissue when exposed to ZnO ENMs, and survival of mussels decrease at an exposure concentration of 2 mg L-1 ZnO ENMs.

These data will be used to build a Dynamic Energy Budget model for mussels

Performance of mussels exposed to ZnO

y = - 0.20x + 0.76R2 = 0.82

NSF: DBI-0830117

Submitted to Chemosphere.

Photoinduced DisaggregationSamuel Bennett, Dongxu Zhou, Arturo Keller

University of California, Santa Barbara

NSF: DBI-0830117

• Natural sunlight and other light sources induce disaggregation of some nanoparticles from the cluster core. This can results in enhanced environmental mobility. We have shown that this enables transdermal penetration of TiO2. We have shown that disaggregation is also observed for CeO2, ZnO and CNTs. Our modeling of the behavior shows that it can be explained theoretically, and that there is a clear explanation for this behavior, which had not previously been observed. The diagram shows how the incident photons increase the energy and allow for partial disaggregation. The graph shows data from our experimental work.

Results to be submitted for publication.

Nanoparticle AggregationDongxu Zhou, Milka Montes, B. Reginald Thio, Arturo Keller

University of California, Santa Barbara

pH 8pH 8, IS = 100 mM

NSF: DBI-0830117

• We are exploring the IRG 1 library for TiO2 NPs with different sizes, shapes and crystal structures. We find that aggregation behavior is a strong function of shape, and that the critical coagulation concentrations differs based on shape. This data will inform the IRG 6 aggregation model.

• Our preliminary studies indicate that the new CEIN NPs (Ag, Pd and Pt) aggregate rapidly in seawater; Ag coated with citrate is stable in freshwater. We are conducting stability studies with alginate, which indicate that stable suspensions can be formed. This information will help IRG 2 & 3 to design their experiments.

Submitted to ACS Nano

Use of a high throughput screening approach coupled with in vivo zebrafish embryo screening to develop hazard ranking for engineered nanomaterials

Saji George1,2, Tian Xia1,2, Robert Rallo2,5, Yan Zhao6, Zhaoxia Ji2, Xiang Wang2, Haiyuan Zhang2, Bryan France3, David Schoenfeld2,6, Robert Damoiseaux3,2, Rong Liu2,5, Shuo Lin6, Kenneth A

Bradley4,2, Yoram Cohen2,5, André E Nel1,21Department of Medicine, Division of NanoMedicine; 2Center for Environmental Implications of Nanotechnology, 3Molecular Shared Screening Resources, 4Department of Microbiology, Immunology and Mol Genetics, 5Chemical and Biomolecular Engineering, 6Department of Molecular,

Cell, and Developmental Biology, University of California, Los Angeles, CA, USA.

We are developing high-throughput screening and in silicodata transformation tools to speed up in vitro hazard ranking of nanomaterials. As a proof of principle, studies were conducted using seven different nanoparticles and eight incremental concentrations and durations of exposure against two cell lines and assayed for four interlinked cytotoxicity events. Using state-of-the-art statistical methods we analyzed, ranked and organized nanomaterials according to cytotoxic potential. Quantum dot (CdSe/ZnS) and ZnO showed the most prominent lethality, Pt, Ag, SiO2, Al2O3 and Au triggered sublethal effects without cytotoxicity. We then compared these results to in vivo response outcomes in zebrafish embryos. Among the results, Ag toxicity in zebrafish differed from in vitro results, which is congruent with this material’s designation as extremely dangerous in the environment.

NSF: DBI-0830117

High throughput screening

Data analysis

In vivo screening using zebrafish embryos

ACS Nano 2011, 5 (3), 1805-1817

Haiyuan Zhang 1*, Tian Xia 2*, Huan Meng 2, Min Xue 3, Saji George 1, 2, Zhaoxia Ji 1, Xiang Wang 1, Rong Liu 4, Meiying Wang 2, Bryan France 6, Robert Rallo 4, Robert Damoiseaux 1, 5, Yoram Cohen 4,

Kenneth A. Bradley 1, 6, Jeffrey I. Zink 3, Andre E. Nel 1, 2, *

1 California NanoSystems Institute, 2 Division of NanoMedicine, Department of Medicine, 3 Department of Chemistry & Biochemistry, 4 Department of Chemical & Biomolecular Engineering,5 Molecular Shared Screening Resources, and 6 Department of Microbiology, Immunology & Molecular

Genetics, University of California, Los Angeles, California, United States

Differential Expression of Syndecan-1 mediates Cationic Nanoparticle Toxicity in Undifferentiated versus Differentiated Normal Human Bronchial Epithelial Cells

Primary cells can provide a more appropriateconnection to in vivo toxicity. We compared theresponse of undifferentiated and differentiatedprimary human bronchial epithelial cells (NHBE) tocationic mesoporous silica nanoparticles (MSNP) thatare coated with polyethyleneimine (PEI) A multi-parametric assay was used to screen for sub-lethaland lethal response outcomes. MSNP coated withhigh molecular weight (10 and 25 kD) polymers weremore toxic in differentiated cells than particles coatedwith shorter length polymers. Differentiated cellsshowed more cellular association with MSNP coatedwith high molecular weight due to more abundantexpression of a proteoglycan, syndecan-1. Thesedata demonstrate the importance of studying cellulardifferentiation as an important variable in theresponse of primary cells to toxic ENM properties.

NSF: DBI-0830117

Undiff Diff

MSNP-PEI MSNP Colocalization of syndecan-1 and MSNP-PEI in differentiated cells

Different cellular association of MSNP-PEI

PEI layer

Differential toxicityACS Nano, 2011, DOI: 10.1021/nn200328m

HTS Data Analysis and Modeling WorkflowYoram Cohen1, Robert Rallo2, Kenneth Bradley1, Andre Nel1, Rong Liu1, Bryan France1,

Robert Damoiseaux1, Saji George1, Haven Liu1

University of California Los Angeles1, Universitat Rovira I Virgili2

Efforts in this project focus on knowledge extraction from high throughput screening data of nanoparticles toxicity, development ofpredictive nano-quantitative-structure-activity relations (nano-SARs), feature selection for nano-SAR development andidentification of pathway linkages. Based on newly developed feature selection approach, a classification based nano-SAR wasdeveloped for cytotoxicity of metal and metal oxide nanoparticles. This work revealed that atomization energy of the metal oxide,period of the nanoparticle metal, nanoparticle volume fraction (in solution), and the primary nanoparticle size were fundamentaldescriptors that enabled correlation of cytotoxicity at high level of accuracy without false negatives. The classification based nano-SAR enables one to identify decision boundaries which are crucial for use in hazard ranking of nanoparticles.

NSF: DBI-0830117

Rallo et. al., ES&T (2011), 45, 1695-1702.

Modeling Transport and Fate of Engineered Nanomaterials (ENMs)Yoram Cohen1, Robert Rallo2, Haven Liu1, Sirikarn Surawanvijit1, Soomin Kim3, Arturo Keller3, Roger Nisbet3

1University of California, Los Angeles, 2Universitat Rovira I Virgili, 3Univerity of California, Santa Barbara

A multimedia modeling scheme is being developed to predict the tranposrt of nanoparticles. The first generation framework was evaluated for the partitioning of TiO2 nanoparticles between air, water and soil, using basic intermeidatransport processes such as wet/dry composition and sedimentation. Quantification of the fate and transport of nanoparticles requires information regarding their size distribution. A predictive computational “constant number” Monte Carlo model ws developed to model NP aggregation and to determine the stable particle size distribution under various environmental conditions. The model performance was successfully tested against experimental DLS data. Implementation of the fate and transport modeling efforts into user-friendly web-based software is an ongoing effort of the Center.

NP input

Microlayer

Atmospheric NP

Resuspension

Sedimentation

AdvectionAggregation

Sediment

DisaggregationWater Body

0

50

100

150

200

250

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0 4 8 12 16 20 24

Hydr

odyn

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m)

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TiO2 pH8 ExperimentTiO2 pH8 SimulationCeO2 pH8 ExperimentCeO2 pH8 Simulation

0

0.5

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0 2000 4000 6000 8000 10000

Stan

dard

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ean

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ove

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(nm

)

Number Of Simulation Particles

Std.Dev<0.5 nm

104 particles

NSF: DBI-0830117

NSF: DBI-0830117 NSF: SES-0531184NSF: SES-0938099

Key findingsneed for regulation, education•Only 46% of participants reported having a nano-specific EHS program. •More (61%) cited “lack of information” as an impediment to implementing nano-specific EHS practices than any other obstacle. •Participants reported high levels of uncertainty about ENM risks•Despite reported lack of information and uncertainty about ENM risk, a majority of participants believes ‘industry knows best’

Engeman, Baumgartner, Carr, Fish, Meyerhofer, Holden, & Harthorn 2011. In Progress.

0%10%20%30%40%50%

Carbon nanotubes

Other carbonaceous

materials

Dry powders Quantum dots Metal Oxides Heavy metals

Almost no risk Slight risk Moderate riskHigh risk Don't know

22% 32% 29%

40%

27%22%

0% 25% 50% 75% 100%

1. It is reasonable to assume that industries working with nanomaterials will adapt or alter their safe-handling

practices when new hazards are discovered.2. Businesses are better informed about their own work-

place safety needs than are government agencies.3. Industries working with nanomaterials can be trusted

to regulate the safe-handling of these materials.4. Voluntary reporting approaches for risk manage-ment are effective for protecting human health and

the environment.5. Employees are ultimately responsible for their own

safety at work.

Strongly agree

AgreeDon’t know

Disagree

Strongly disagree

International Survey of EH&S in Nanomaterials Industry

Environmental Sociology Case StudiesMary Collins1, William Freudenburg1, Barbara Herr Harthorn1, Terre Satterfield2

1University of California, Santa Barbara, 2University of British Columbia

Nanoremediation: Will equity concerns arise?Collins, Harthorn (UCSB), Satterfield (UBC)Nanoremediation currently in use in 50 US sites--in situ mitigation rather than removal • Benefit or risk, dep. on safety and effectiveness• Is there balance in subpopulation distribution of nanoremediation siting? Spatial analysis• Preliminary results indicate likely balance, but…

Temporal Aspects of Public-Private PartnershipsCollins, Freudenburg (UCSB) •Relationships that are Unproblematic initially may become problematic in thecontext of risk management •Implication: Use caution in forming industry partnerships that could lead toa perception of compromised risk management Nanotechnology and RecreancyFreudenburg, Collins (UCSB) •Theoretical contribution addresses potential risks to safe nanotech, reminding CEIN to instill public trust through its own safety practices, and transparent and timely disclosure of risks, in accessible language(Expected Publication: 2011 Social Life of Nanotechnology eds: Harthorn & Mohr)

NSF: DBI-0830117

Nanoecotoxicology Lecture SeriesHilary Godwin

UC CEIN Education/Outreach Director

Each lecture in this 13 lecture series includes learning objectives, required and recommended readings, and quizzes, which can all be accessed through a password-protected website. This lecture series is available to Center members and to external partners, like CINVESTAV and INSP in Mexico, interested in building research programs in this area.

As a result of this lecture series, UC-CEIN will host a “Nanoecotoxicology Bootcamp” at UCLA in August 2011 to provide capacity building for researchers in Mexico and to promote collaborations between women scientists in the United States and women scientists in Mexico.

Postdoctoral Fellow Saji George explains High Throughput Screening (HTS) to visitors at the UC-CEIN’s HTS facility. Through the Nanoecotoxicology online lecture series, researchers unable to visit Los Angeles can listen to Dr. George’s lecture.

NSF: DBI-0830117

Public OutreachHilary Godwin

UC CEIN Education/Outreach Director

Partnerships with university-based and community-based organizations have resulted in public outreach events that focus on communicating key scientific concepts to the community. UC-CEIN Volunteer Educators lead interactive tabletop activities and engage the public in dialogue about nanoscale science and engineering.

Event Location Attendees CEIN Volunteers

NanoDays 2010, Los Angeles California ScienCenter 550 14

NanoDays 2010, Santa Barbara SB Museum of Natural History 500 4

Nanotechnology: Small is Big! Santa Monica Public Library 50 5

Small things from a big planet SB Public Library 50 3

SciArt summer camp UCLA 50 10

Explore your universe! UCLA 500 20

NSF: DBI-0830117


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8. Strategic Research Plan. Long‐term Research Goals of the UC CEIN A long‐term goal of the UC CEIN is to establish a series of broad‐based predictive and integrative scientific approaches in which nanomaterial bio‐physicochemical interactions at cellular, subcellular and organism levels are utilized for prioritizing in vivo eco‐toxicity testing to predict how engineered nanomaterials (ENMs) may impact a variety of biological lifeforms at different trophic levels in the environment. Through the establishment of a rigorous scientific platform that attempts to link (i) nanomaterial physicochemical properties to (ii) mechanisms and pathways of injury at cellular, organism and population levels, (iv) high throughput screening and in silico data transformation, (v) fate and transport, (vi) exposure, (vii) bioavailability and trophic transfer, our goal is to establish a scientific paradigm on which to base assessments of the potential environemtnal impact of ENMs. In order to achieve these long‐term goals, it is necessary to establish well characterized compositional and combinatorial nanomaterial libraries that will allow us to determine how key physicochemical properties such as chemical composition, size, shape, aspect ratio, porosity, solubility, crystalline states, electronic and redox potential, band gap, surface charge, purity and particle dispersal or aggregation state determine material distribution and compartmentalization within the environment, cellular uptake, bioavailability and catalysis of bio‐catalytic activity that could lead to toxicity in bacteria, algae, phytoplankton, protozoa, mammalian cells and select sets of trophic life forms in terrestrial and aquatic ecosystems. In order to develop in vitro toxicological testing that will inform in vivo testing and provide information that is necessary for guiding the decision making process regarding ENM's impact on the environment. In this regard, an important goal is to develop high content and high throughput screening that can be used to rapidly screen the ENM libraries as well as demonstrate how the systematic physicochemical variation of select material properties can influence biocompatibility or biotoxicity. The number of materials that can be handled simultaneously and the volume of data generation through high throughput screening are important for establishing high content data sets as well as for meeting the scale of observations that is required to cover a wide range of nanomaterials. An important approach within the Center to prioritize research towards critical ENM and organismal interactions has been to use a series of microcosms using terrestrial, fresh water and marine organisms at different trophic levels to determine whether the in vitro and high throughput testing is predictive of the toxicological outcomes in more complex environments. As an example of this interfacing, we are using zebrafish embryo screening to check the in vivo relevance of high throughput data collected in mammalian and fish cells. Similar connectivity is also being sought across the Center in studies looking at bacteria as basal food web organisms and protozoa as feeders, hematocytes and sea urchin embryos, phytoplankton as primary producers and copepods as feeders etc. Moreover, dynamic energy budgeting is being used as an important integrative model in population studies. These microcosm studies also serve to understand and predict the mobility and bioavailability of the nanomaterials in complex environmental matrices.

An important long‐term objective is to utilize the structure‐activity relationship of nanomaterial physicochemical properties and other suitable descriptors with in vitro and eco‐toxicological outcomes to develop guidelines for safe design of engineered nanomaterials as well as methods that can be implemented to lessen the impact of hazardous principles in the environment. As an example, we are using heat maps, self‐organizing maps and feature selection approaches to identify metal and metal


33

oxide NP properties and descriptors (e.g., band gap, periodicity, and solubility that can be further modified by size, state of aggregation, shapte) to develop a better understanding of the association between cytotoxicity and oxidative stress. Another key UC CEIN goal is to use the knowledge generation through integrated research activities and risk perception surveys to inform the public, academia, industry and government agencies how nanotechnology can be safely implemented in society and the marketplace. By reducing uncertainty about potential nanomaterials toxicity, our goal is to promote widespread acceptance of nanotechnology in society. Organization and Integration of Center Research Activities Our predictive and integrative scientific models will consider the nanomaterials most likely to come into contact with the environment. In the first five years of our research activities, we are focusing extensively on metal and metal oxide nanomaterials as well as single‐wall and multi‐wall carbon nanotubes (CNT). We are considering the physicochemical properties of these materials that allow them to spread to the environment, their bio‐availability through cellular/organismal uptake and ability to perform biocatalytic activities that could lead to toxicity in bacteria, algae, phytoplankton, protozoa, mammalian cells and select sets of trophic lifeforms in terrestrial, fresh water and marine ecosystems. We are considering mechanisms and biological pathways of injury that can be used to perform high content and high throughput screening with a view to facilitate in vivo toxicological assessment, including development of cost‐effective and rapid screening paradigms. All of the above nanomaterial physicochemical characteristics, and biological and toxicological data are being used to establish in silico decision making tools that can help to perform hazard ranking, establish structure‐activity relationships and through safe‐by‐design approaches to implement risk reduction strategies.

Our research goal of developing a series of predictive risk models for nanomaterial impact on the environment is currently being conducted by seven IRGs. To develop an understanding of the QSARs, IRG 1 is acquiring a physical library of standard reference nanomaterials representing the major classes of representative nanomaterials from commercial sources or in‐house synthesis. Nanochemistry experts in IRG 1 use advanced NM design and synthesis methods to develop compositional and combinatorial libraries that consist of a single

material made in different sizes and shapes or with different dissolution, band gap distribution, surface charge, and crystalline states. These nanomaterials are being characterized to determine the physicochemical properties (IRG 1) that are associated with cellular, tissue, and systemic injury in a variety of environmental life forms (IRG 2 and 3). These ecological life forms are chosen to represent potential sentinel species for ecotoxicological testing as well as different trophic levels in fresh water, seawater and terrestrial environments that can be used for studying exposure, bioaccumulation, trophic transfer and dynamic energy budget modeling (IRG 3). The engineered NPs are also being evaluated to


34

determine their state of aggregation, stability, and transport in various environmental aquatic and tissue culture media (IRG 4). We use the key interfacial properties governing interactions at the nano–bio interface (size, surface area, shape, aggregation, dispersal, charge, dissolution) to develop HTS approaches (IRGs 1 & 5) allowing contemporaneous testing of batches of nanomaterials in representative cellular systems (e.g., bacteria, mammalian cells) for hazard prediction based on final common toxicological pathways (oxidant stress response, proliferation, ATP production, mitochondrial dysfunction, apoptosis). In the case of the metal and metal oxides we are using information about cellular toxicity together with nanomaterial descriptors to develop quantitative structure‐activity relationships (QSARs) to assist hazard ranking of ENMs (IRG 5 and 6). In order to develop tools for decision analysis based on in silico toxicity information, fate and transport data models, and other pertinent information that would enable hazard raking for ENMs, we are developing software tools that rely on both qualitative and quantitative information from across the Center (IRG 6). The above developments interface with IRG7 which is conducting risk perception surveys among the public and industry in order to shape our research and provide environmental safety guidelines to regulatory agencies (IRG 7). 5 Year Milestones


35

9. Research Program, Accomplishments, and Plans The UC CEIN has successfully integrated the expertise of engineers, chemists, colloid and material scientists, ecologists, marine biologists, cell biologists, bacteriologists, toxicologists, computer scientists, biostatisticians and social scientists necessary to create a predictive scientific platform to inform us about the possible hazards and safe design of nanomaterials (NMs) that may come into contact with the environment. Now in our third year of operation, there are 38 distinct but interactive research projects active across 7 IRGs, along with 3 major service cores (ENM production, characterization and distribution in IRG 1; Molecular Shared Screening Resource for HTS analysis in IRG 5; and Data Management/Collaborative Infrastructure in IRG 6). The Center is organized into 7 interdisciplinary research groups (IRGs):

IRG 1: Nanomaterial Standard Reference and Combinatorial Libraries and Physical‐Chemical Characterization.

IRG 2: Studying Nanomaterials Interactions at the Molecular, Cellular, Organ, and System Levels

IRG 3: Organismal and Community Exotoxicology

IRG 4: Nanoparticle Fate and Transport

IRG 5: High‐Throughput Screening (HTS), Data Mining, and Quantitative‐Structure Relationships for Nanomaterial Properties and Nanotoxicity

IRG 6: Modeling of the Environmental Multimedia Nanomaterial Distribution and Toxicity

IRG 7: Risk Perception of Potential Environmental Impacts of Nanotechnology For each IRG, the goals, organization and integration, major accomplishments, and plans for the coming year are presented in detail in the following pages. Detailed information about each research project, including an abstract of the project, are available on the CEIN website: http://www.cein.ucla.edu Seed Funding In Winter 2010, the UC CEIN Executive Committee issued our second call for seed funding proposals. The seed funding competition is designed to support new integrated research projects that cannot be carried out in the scope of existing project funding. Innovative and cross‐cutting proposals were sought that could be completed or can show definitive progress within a year of funding. Proposals were limited to existing CEIN faculty members. The executive committee reviewed all proposals and selected 4 seed proposals for funding effective June 1, 2010 for a period of 1 year. This year's seed projects funded are:

IRG 1‐11: Trojan Horse Nanoparticles ‐ Jeff Zink ‐ UCLA

IRG 2‐8: High Throughput Screening Sublethal Effects of Engineered Nanomaterials to Bacteria ‐ Trish Holden ‐ UCSB, Ken Bradley ‐ UCLA

IRG 2‐10: Integrated High Throughput Screening for Marine and Estuarine Phytoplankton and Primary Production ‐ Hunter Lenihan, Arturo Keller, Roger Nisbet ‐ UCSB, Gary Cherr ‐ UC Davis (incorporated into IRG 3‐1)

IRG 5‐6: Nanotoxicity Testing in Zebrafish ‐ Andre Nel, Tian Xia, Jeffrey Zink ‐ UCLA, Gary Cherr ‐ UC Davis

In April/May 2011 the Executive Committee will review the progress and success of introducing new integrative topics into the Center portfolio. It is fully expected that all four seed projects will remain active in our portfolio. A new call for proposals for the Year 3 seed funding program will be issued and selected projects will begin in May/June 2011.


36

IRG 1 – Nanomaterial Standard Reference and Combinatorial Libraries and Physical‐Chemical Characterization

Faculty Investigators: Carolyn Bertozzi, UC Berkeley/Lawrence Berkeley Lab – Professor, Chemitry, Molecular/Cell Biology Freddy Boey, Nanyang Technological University – Professor, Materials Science Engineering Jeffrey Brinker, University of New Mexico/Sandia – Professor, Chemical/Nuclear Engineering Robert Haddon, UC Riverside – Professor, Chemistry Mark Hersam, Northwestern University ‐ Professor, Materials Science & Engineering Joachim Loo, Nanyang Technological University – Assistant Professor,Materials Engineering Lutz Madler, University of Bremen – Professor, Materials Science Andre Nel, UCLA – Professor, Medicine; Chief, Division of NanoMedicine Galen Stucky, UC Santa Barbara – Professor, Chemistry and Biochemistry Jeffrey Zink, UCLA – Professor, Chemistry and Biochemistry – AREA LEAD Number of Graduate Students: 8 Number of Undergraduate Students: 1 Number of Postdoctoral Researchers: 3 Goals of IRG 1: IRG 1’s main goals are to assemble nanomaterial composition and combinatorial libraries, to characterize the physiochemical properties of the nanomaterials, to synthesize “designer” or “hand‐crafted” specialty nanomaterials and characterize them, and to identify new and important nanomaterials that could or should be of interest for environmental impacts. Organization and Integration of IRG 1 Projects Currently IRG 1 includes a core dedicated to nanomaterial library acquisition, distribution and characterization and 9 ongoing research projects: Core Function:

IRG 1‐1: Managing library production, distribution, and characterization (Tables 1‐4) (Jeffrey Zink)

Current IRG 1 Research Projects:

IRG 1‐4: Ag Nanoparticle Control and Bioprocesses and Cytotoxicity (Galen Stucky)

IRG 1‐5: Processing and Characterization of Single Walled and Multi Walled Carnon Nanotubes (Mark Hersam). A new direction involving commercial SWNTs prepared by three different methods has been introduced. Prof. Mark Hersam (Northwestern University) was recruited for this project; his group separates purified SWNTs from the raw commercial mixtures.

IRG 1‐6: Metal Oxide Nanoparticle Library – Labeling and Synthesis for Toxicology and High Throughput Testing in the CEIN (Jeffrey Zink)

IRG 1‐7: FSP Generated Pure and Fe Doped ZnO or TiO2 NP Libraries for Testing Paradigms of Environmental and Cellular Responses (Lutz Madler)

IRG 1‐9: Developing a Database of Nanomaterials with Potential Biomedical and Industrial Applications Correlating Their Physiochemical Properties to Toxicity (Joaquim Loo, Singapore)

AveryCEIN

Underline


37

IRG 1‐10: Systematic Synthesis of Nanoparticles of controllable morphology, composition and porosity to perform biological structure‐function analysis in mammalian cells and bacteria (Jeff Brinker)

IRG 1‐11 Trojan Horse Nanoparticles (Seed Project) Jeffrey Zink, Andre Nel, Trish Holden The organization of IRG 1 involves synthesis of nanoparticles at UCLA (IRGs 1‐6 and 1‐11), at UCSB (IRG 1‐4) at Sandia National Laboratory (IRG 1‐10) and at the University of Bremen (IRG 1‐7). Physical and chemical characterization of the materials are partially carried out in these laboratories, and final and full characterization, cataloging, warehousing and distribution are carried out by IRG 1 at UCLA. Integration is coordinated through IRG 1. By mutual agreement reached by teleconferencing, each research group is focused on a specific type of nanomaterial as indicated by the titles of the projects. Commercially available materials are purchased primarily by IRG 1‐1. The IRGs are all in frequent communication with Dr. Zink and IRG 1‐1. Major Accomplishments of IRG 1 (since March 2010) At the beginning of calendar year 2010, IRG 1 had completed its initial major goal of assembling and characterizing a library of standard reference materials. Characterization of the three commercial nanoparticle compositions (TiO2, ZnO and CeO2) and several new compositions was completed. Particles were distributed to the high throughput screening group and have also been distributed to all of the biology groups that requested them (Tables 5‐6). The library was expanded in 2010‐11, primarily by including a wide range of sizes and shapes of titania, ceria and silica nanoparticles. The updated library is summarized in Table 1. In addition, planning of the next generation of library materials was discussed extensively among the IRG group leaders, and as a result of those discussions, five new types of materials were added to the library. The materials (and associated IRG subgroups) are:

1. Silver nanoparticles (IRG 1‐8). The first set of materials were commercially‐synthesized 20 nm and 40 nm particles with citrate coatings. Monodispersed particles with 10 nm diameters and polydispersed particles with an average diameter of 57 nm were subsequently added.

2. Single wall carbon nanotubes (IRG 1‐5). Samples of as‐synthesized and of purified materials (1.5 gm each) from three different commercial sources prepared by three different methods were purchased and used as received. The Hersam group purified smaller amounts (100 mg) of the samples and provided tubes of uniform diameters for microcosm studies.

3. Ceria nanoparticles (IRG 1‐1 & 1‐6). Because of differences in gram negative and gram positive bacterial responses, ceria has been selected for more detailed studies. Samples of different shapes (cubes, rods, wires) have been synthesized using hydrothermal methods.

4. Pt nanoparticles. Commercial Pt nanoparticles were purchased, characterized and added to the library.

5. Semiconductor (3‐5) nanoparticles such as GaAs. Because of their use in photovoltaic devices, these particles are of interest. Decisions about the composition, size and coatings are currently being discussed.

Postdoctoral fellow Raffaella Buonsanti is working at the Molecular Foundry on metal oxide and Pt nanoparticles. A library of aluminum‐doped titanium dioxide with different sizes, shapes (pyramids, spheres, rods) was developed; automated synthesis will begin when the Foundry’s automated robotic system is back on line in March 2011. These particles have a populated conduction band and will be used to study toxicity caused by electron transfer from the particles and superoxide production.


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Characterization of new and designer materials is continuing actively. Metal nanoparticles, designer metal oxides, and doped metal oxides are currently being examined. IRGs 1‐1 and 1‐6 expanded their characterization capabilities to include surface area and pore volume measurements by using gas absorption/desorption isotherms and BET analysis. A high throughput instrument that allows four simultaneous measurements to be performed was purchased and is currently being used in the CEIN laboratories to study the surface areas and pore volumes of library materials. Fluorescent labeling of nanoparticles in the above libraries has been carried out at the request of individual research groups. Important results are reported in the summaries of IRGs 2 and 5. In addition, titania nanoparticles were coated with cationic and anionic molecules and the particles distributed to IRGs 2 and 5 for testing the hypothesis that cationic particles are more toxic than anionic ones. Extensive studies of metal ion doping (IRG 1‐7) on ZnO toxicity have been carried out. Iron doping reduced the dissolution rate. Iron‐doped titania was also made and characterized. Shifting the band gap energy into the UVA region of the spectrum enabled studies of photo‐toxicity caused by electron hole pairs in the nanomaterial without direct deleterious effects of the light on the cells. Crystalline silica nanoparticles have been successfully synthesized and current work is aimed at scaling up the fabrication of highly monodisperse crystalline silica. Both silicalite (IRG1‐6) and quartz samples (IRG 1‐10), are undergoing high thoughput analysis to probe the role of crystallinity (faces, sharp edges, strained rings) on toxicity. Commercial silica (including fumed silica that appears to be toxic in the preliminary high throughput studies) was purchased and characterized. Stealth or Trojan horse nanoparticles containing copper oxide, iron oxide and silver cores were synthesized (IRG 1‐11). Empty Trojan horse nanoparticles (without a metal or metal oxide core) were made and given to IRG 2 for initial study. Tetrahymena thermophilia ingest these particles extensively with no toxicity. Detailed studies are in progress. Impacts on the Overall Goals of the Center The major impacts on the overall goals of the center directly follow from the major accomplishments. Assembly of nanoparticle library containing over 100 different types of particles (condensed version in Table 1) was necessary in order to provide materials for high throughput screening. Characterization of all of the nanoparticles in the current library (methods in Table 2 and condensed results in Table 3) was necessary in order for the researchers to know exactly what physical and chemical properties the materials possess. Identification of efficient and reliable dispersion methods was a major step forward in helping the researchers prepare the samples in stable suspended forms for the biological studies. Samples prepared and characterized by IRG1 are currently being used throughout the entire center. The distributions of samples (silica and silver as representative examples) to individual research groups are summarized in tables 5 and 6. Major Planned Activities for the Next Year: The structure of IRG1 has been reorganized in response to the evolving needs and emphasis of the CEIN. One of the major changes involves IRG1‐1 that will in future be regarded as a core function rather than a project. An extensive library of nanomaterials has already been assembled, and supplies of


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nanoparticles, characterization details about the materials, and methodology for enhancing the suspendability of the particles in biologically relevant solutions have been well established. In order to increase the interactions between the creators of the library and the scientists carrying out studies with organisms, the current staff research scientist will take on a more formal role as the liaison between this IRG1 core and the other IRGs. An additional staff scientist will be hired to carry out the continuing acquisition, characterization and distribution duties to free up Dr. Ji’s time for the coordination activities. Interest in carbon nanotubes is high due to the increasing industrial importance of these carbonaceous materials. The role of the current IRG1‐6 will be enhanced provided that there is suitable need for and use of CNTs in ecotoxicity research. There is continuing interest in metal oxide nanoparticles and the role of their valence and conduction band energies on cellular and organismal responses, and in pure metal nanoparticles and the role of the crystalline faces, surface defects, and dissolution on environmental impacts. There is increasing recognition that surface coatings play a major role in both dispersability of particles, transport of the particles in the environment, and toxological responses. Research efforts involving the capping agents used in the particle syntheses, induced surface properties (hydrophilicity/hydrophobicity, sign of surface charge) caused by molecules on the surfaces, nature of fluorescent tags needed for monitoring, and effects of molecules adsorbed on surfaces (both deliberately for dispersion and adventitiously from the surrounding medium) will be increased. Concluding Observations The researchers in IRG 1 have accomplished their initial goal of assembling and carefully characterizing a library of commercially available standard reference materials and of materials synthesized by members of the IRG and making them available to the members of the CEIN. The acquisition and characterization of the second generation of library materials as determined by the IRG leaders is almost complete. New libraries of “designer” nanoparticles are rapidly expanding, especially those involving doped metal oxide particles, metal nanoparticles, multiple forms of silica particles, and carbon nanotubes.


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Table 1. Nanoparticle Library in the CEIN

Composition Size (nm) Shape Phase/Structure

TiO2

~25 Spheres 80% Anatase & 20% Rutile

6, 10, 15, 40, 60, 100, 260 Spheres Anatase

5, 12, 60, 140 Spheres Rutile

4×15, 8×45, 12×60, 10×100, 30×100, 80×300

Rods Rutile

5×1000 Wires Rutile

5, 10, 15, 20, 30, 60, 130, 160, 220, 240, 460, 600

Spheres Amorphous

15×70 Rods Amorphous

CeO2

5, 7, 10‐12, 15‐30, 20‐70 Cubes Crystalline

8×33, 7×39, 7×51, 7×75, 7×94, 12×500, 12×1000‐2000

Rods Crystalline

ZnO ~20 Spheres Crystalline

Fe‐ZnO 20, 15, 14, 12, 8, 8, 8 nm with 0, 1, 2, 4, 6, 8, 10 atomic weight% Fe

Spheres Crystalline

SiO2

5, 8, 30, 50, 80, 130 (Commercial) Spheres Amorphous

6, 30, 40, 60, 70, 135, 200, 500, 600, 2000 (In‐House)

Spheres Amorphous

81×137, 94×209, 72×201, 65×308, 69×446 Rods Mesoporous

Ag 8, 21, 40 (Monodispersed); 57 (Polydispersed)

Spheres Crystalline

Pd 13 Spheres Crystalline

Pt 5‐10 Spheres Crystalline

SWNT

~3×1500 (Caboxylated) Nanotubes Crystalline

0.7‐0.9×450‐2000 (Purified) Nanotubes Crystalline

1.5×1000‐5000 (As‐prepared, Purified) Nanotubes Crystalline

0.8‐1.2x100‐1000 (As‐prepared, Purified) Nanotubes Crystalline

MWNT

2‐6 nm inner diameter, 5‐20 nm outer diameter, 1‐10 µm length (As‐prepared)

Nanotubes Crystalline

2‐6 nm inner diameter, 5‐20 nm outer diameter, 1‐10 µm length (PEI‐coated)

Nanotubes Crystalline

68±20 nm outer diameter, up to 300‐400 µm length (Aligned) Nanotubes Crystalline


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Table 2. Overview of Analytical Methods for Physical and Chemical Characterization Transmission Electron Microscopy (TEM) & Scanning Electron Microscopy (SEM):

The operation principles of electron microscopy are based on the interaction between an electron beam and a solid surface. In TEM, transmitted or forward‐scattered electrons are used to obtain images; while in SEM, back scattered or secondary electrons are analyzed to yield images. With enough representative images, TEM and SEM can be used to obtain primary size, morphology, topography, state of agglomeration, or even some crystallographic information of nanoparticles.

X‐ray Diffraction (XRD):

When a coherent X‐ray beam is directed at a sample, interaction of the X‐rays with the sample creates diffracted beams that can be related to interplanar spacings in the crystalline sample according to the Bragg’s law: nλ=2dsinθ; where n is an integer, λ is the wavelength of the X‐rays, d is the interplanar spacing, and θ is the diffraction angle. Based on this principle, XRD can be used to identify crystalline phase and structure and to determine crystallinity. Primary size of nanoparticles can also be derived from the XRD patterns using the Sherrer equation: S=λ/ωcosθ; where S is the particle size and w is full‐width‐at‐half‐maximum of the diffraction peak.

Dynamic Light Scattering (DLS):

In DLS, a monochromatic light beam is directed at a particle suspension where it is scattered. Due to the random Brownian motion of the particles, the intensity of the scattered light fluctuates with time; from which a translational diffusion coefficient, Dt, can be determined using an autocorrelation function. Hydrodynamic diameter (dH) of nanoparticles can then be estimated from the Stokes‐Einstein equation dH=kT/3πηDt; where k is the Boltzmann constant, T is the temperature, and η is the viscosity. Information including size distribution and state of agglomeration can also be derived from the DLS measurement.

Zeta Potential & Electrophoretic Mobility (EPM):

Surface charge can be determined indirectly by measuring the zeta potential (ζ) or electrophoretic mobility (EPM) of the particles. The EPM is defined as the velocity of a particle per electric field unit and is obtained by applying an electric field to the particle suspension and measuring the average velocity of the particles. Using Smoluchowski or Huckel equation, the zeta potential can be calculated. The magnitude of zeta potential gives an indication of the potential stability of the nanoparticle suspension. Nanoparticles with zeta potential more positive than 30 mV or more negative than ‐30 mV are normally considered stable. By adjusting pH, an isoelectric point (pHiep) can also be determined. Nanoparticles are positively charged below pHiep and negatively charged above pHiep.

Gas Absorption/Desorption, Brunauer‐Emmett‐Teller (BET) Surface Area Analysis:

BET method is based on adsorption of gas on a surface. The amount of gas adsorbed at a given pressure is used to determine the specific surface area. Assuming the particles have solid and uniform spherical shape with smooth surface, an average particle size can also be estimated.

Thermo‐Gravimetric Analysis (TGA):

TGA is usually used to determine a material’s thermal stability and its fraction of volatile components by monitoring the weight change as a function of temperature and time. Based on the weight loss or gain profile, kinetic process such as dehydration, oxidation and decomposition can be identified and the corresponding moisture content and organic content can be determined.


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Cross Polarization/Magic Angle Spinning Solid State NMR (CP/MAS SSNMR)

This technique provides molecular information about solid samples similar to that obtained from conventional solution phase nmr. The attachment of linkers is observed by using 29Si, and information about the attached surface functional groups and molecular machines is obtained from 13C and 1H nmr.

UV/Visible Absorption Spectroscopy:

Absorption spectroscopy provides information about molecules in pores or attached on the surface of particles. It is especially useful for monitoring attachment of molecules with chromophores that have large molar absorptivities in the ultraviolet or visible regions of the electronic spectrum. The technique can also be used for nanoparticle suspension stability evaluation. In this case, the absorbance at a characteristic wavelength, which can be linearly correlated to the nanoparticle concentration, will be monitored as a function of time.

Fourier Transform Infrared Spectroscopy (FTIR)

Infrared spectroscopy spectroscopy provides information about molecules in pores or attached on the surface of particles. It is especially useful for monitoring attachment of molecules that have vibrational absorption bands in regions that are not obscured by absorptions of the particle itself.

X‐ray Photoelectron Spectroscopy (XPS) In XPS the sample is illuminated with soft x‐ray in an ultrahigh vacuum, which leads to the production of photoelectrons from the sample surface. From energy analysis of the generated electrons, the binding energy, which is characteristic of each element, can be calculated. The binding energy of a particular electron is also affected by its surrounding environment, therefore can also provide oxidation state information. Peak area of each element allow for quantitative analysis. In the case of analysis for nanoparticles, defect information can also be obtained.

Inductively coupled plasma mass spectroscopy (ICP‐MS) ICP‐MS employs a plasma (ICP) as the ionization source and a mass spectrometer (MS) analyzer to detect the ions produced. The instrument can perform multi‐elemental analysis with high sample throughput and excellent sensitivity. It can determine analyte concentration down to the part per trillion level (ppt). It is one of the most powerful techniques for elemental analysis and purity quantification in nanomaterials.

Flame Photometry Flame photometry is used for determination of certain metal ions such as sodium, potassium, and calcium. It relies on the principle that an alkali metal salt drawn into a non‐luminous flame will ionize, absorb energy from the flame and then emit light of a characteristic wavelength as the excited atoms decay to the unexcited ground state. The intensity of emission is proportional to the concentration of the element in the solution. Therefore it is suitable for both qualitative and quantitative determination.


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Table 3. Physical and chemical characterization of selected nanoparticles added to the CEIN nanomaterial library in 2010‐11.

SRM Properties Technique Unit Ag Pd Pt CeO2 CeO2 SG 65‐SWNT

P2‐SWNT

Primary Size TEM nm 21±3 57±20 5‐10 8 × 33 7 × 94 0.7‐0.9 × 450‐2000

1.4 × 500‐1500

Shape TEM Sphere Sphere Sphere Rods Rods Wires Wires

Hydrodynamic Diameter

DLS nm 29.9±0.5 106±7 133±10 267±9 336±5 458±128 664±206

Phase & Structure

XRD Crystalline Crystalline Crystalline Crystalline Crystalline Crystalline Crystalline

ζ in DI water (50 µg mL‐1)

ZetaPALS mV ‐36.6±0.9 ‐36.0±2.0 ‐50.7±3.0 40.6±1.6 36.0±1.3 ‐23.4±1.0 ‐29.3±2.4

EPM in DI water (50 µg mL‐1)

ZetaPALS 10‐8 m2 V‐1 s‐1

‐2.75±0.10 ‐2.47±0.11 ‐3.82±0.23 3.05±0.12 2.71±0.10 ‐1.76±0.08 ‐2.21±0.18

Table 4. Nanoparticles in the process of being characterized.

Composition Estimated Size (nm) Other Properties/Information

Ag 10 1mg/mL monodispersed spheres

10 Cysteine‐coated spheres, powder

CuO 10

Highly dispersed aqueous suspension made by hydrothermal synthesis

120 × 60 × 500 (Width × Height × Length) Nanorods made by hydrothermal synthesis

SWNTs

0.7‐0.9 × 450‐2000 Well dispersed nanotubes in 1% w/v F108NF Pluronic aqueous solution

1.4 × 500‐1500 Well dispersed nanotubes in 1% w/v F108NF Pluronic aqueous solution

0.8‐1.2 × 100‐1000 Well dispersed nanotubes in 1% w/v F108NF Pluronic aqueous solution


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Table 5. SiO2 Nanoparticle Distribution in UC CEIN (2010‐11).

Sample Name Size (nm)

Quantity (g)

Requestor IRG # Shipping Date

Commercial 5 nm colloidal SO2 5 0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010



Commercial 80 nm colloidal SiO2 80 0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010

Commercial 130 nm colloidal SO2 130 3.6 Robert Miller 3 3/23/2010

0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010

AML silica #2 6 0.3 Robert Miller 3 3/23/2010


AML Aug 6 2010‐2 silica 8 8 0.01 Sijie Lin 5 9/9/2010

AML silica colloid July 7 #2 29

0.45 Robert Miller 3 3/23/2010

0.03 Angela Ivask 5 3/24/2010


AML silica #4 32 0.01 Sijie Lin 5 9/9/2010

AML Mar 31B Silica 65 10 Reginald Thio 4 5/4/2010

5 Reginald Thio 4 6/11/2010

AML Aug 6 2010‐5 silica 65 10.8 Reginald Thio 4 8/20/2010

AML silica colloid July 2 #1 72

0.49 Robert Miller 3 3/23/2010


0.01 Sijie Lin 5 9/9/2010

AML Mar 31A Silica 135 0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010

AML April 1C Silica 200

9 Reginald Thio 4 5/4/2010


0.01 Sijie Lin 5 9/9/2010

AML Aug 6 2010‐6 silica 200 10 Reginald Thio 4 8/20/2010

AML Mar 30B Silica 530 0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010

0.01 Sijie Lin 5 9/9/2010

AML Mar 30A Silica 640 10 Reginald Thio 4 5/4/2010

AML Mar 31C Silica 2000 0.02 Haiyuan Zhang & Saji George 2 & 5 5/12/2010

0.01 Sijie Lin 5 9/9/2010


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Table 6. Silver Nanoparticle Distribution in UC CEIN.

Sample Name Size (nm)

Quantity (g)

Requestor IRG # Shipping Date

Silver NP Powder (Dr. Hoek) ~20

0.5 Gary Cherr & Arturo Keller 3 & 4 3/19/2009

1 Gary Cherr & Arturo Keller 3 & 4 3/19/2009

0.5 Angela Ivask 5 3/26/2011

Silver NP Powder (Sigma Aldrich) 57 0.05 Dongxu Zhou 4 11/12/2010

0.25 Angela Ivask 5 11/23/2010

20 nm BioPure Ag Nanoparticles 21

0.03 Reginald Thio 4 10/5/2010

0.001 Saji George 2 11/1/2010

0.0015 Angela Ivask 5 11/23/2010

0.006 Angela Ivask 5 12/8/2010

40 nm BioPure Ag Nanoparticles 40

0.03 Reginald Thio 4 10/5/2010

0.001 Saji George 2 11/1/2010

0.0015 Angela Ivask 5 11/23/2010

0.006 Angela Ivask 5 12/8/2010


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IRG 2: Studying NMs’ Interactions at the Molecular, Cellular, Organ, and Systemic Levels Faculty Investigators: Gary Cherr, UC Davis – Professor, Environmental Toxicology/Nutrition Patricia Holden, UC Santa Barbara – Professor, Environmental Microbiology – AREA LEAD André Nel, UCLA – Professor, Medicine; Chief, Division of NanoMedicine Roger Nisbet, UC Santa Barbara – Professor, Ecology, Evolution, Marine Biology Galen Stucky, UC Santa Barbara – Professor, Chemistry and Biochemistry Number of Graduate Students: 6 Number of Postdoctoral Researchers: 8 Goals of IRG 2: There are six major goals for IRG 2: 1. Explore the mechanisms of NP uptake into cells, tissues, and organs. Mechanisms include: electron

transfer from membranes and cell components to NPs, ROS production and membrane or organelle damage from energized NPs, NP uptake via various mechanisms including membrane wrapping and phagocytosis, and trophic transfer. These mechanisms will vary between eukaryotic and prokaryotic cell systems.

2. Study paradigms for toxicity that can be used to screen for the potential adverse environmental

impacts of NPs. Paradigms include: cellular oxidation via electron transfer from cells or organelles to NPs, toxic metal release from metallic NPs—either extracellularly with passive uptake of ions or intracellularly where NPs effectively deliver large doses released upon NP breakdown, sorption of ambient toxicants onto NPs and delivery into cells with NP uptake, intracellular ROS‐mediated oxidative damage, lysosomal destabilization or mitochondrial function impairment (both potentially leading to mammalian cellular apoptosis), cellular oxidative stress response induction, direct targeting of NPs to specific organelles, energy‐dependent expulsion of NPs, intracellular dissolution into toxic constituents, and extracellular binding leading to nutrient deprivation. As above, these paradigms will vary with bacterial or mammalian cell systems.

3. Evaluate localization of NPs in cells and tissues. Of interest are membrane binding as a perquisite to

either uptake or toxicity, receptor roles, organelle targeting, NP intracellular integrity, methods to evaluate the “aging” of NPs once inside cells and tissues, and roles of NP characteristics including capping agents.

4. Evaluate whole organism responses including developmental stages (embryonic) to determine:

developmental effects, whole organism systemic functions (immune, digestive, respiratory), and NP localization, stability and bioprocessing.

5. Evaluate population responses including changes in respiration, growth rates and extents,

reproduction, viability, and NP uptake and modification. 6. Model effects using DEB theory as a framework which employs submodels in toxicokinetics and toxic

effects.


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Organization and Integration of IRG 2 Projects: There are currently 9 projects within IRG 2. The organization is such that IRG2‐1 and IRG2‐7 are being performed at UCLA in the Nel Lab and in the MSSR (high throughput screening or HTS) facility; IRG2‐2 is being performed at UCD/BML in the Cherr Lab; IRG2‐3 is being performed at UCSB in the Holden Lab; IRG2‐4 is being performed at UCSB in the Nisbet Lab; IRG2‐5 is being performed at UCSB in the Holden Lab with the Cherr Lab (UCD/BML) and with IRG1 including the Stucky Lab (UCSB) and the Zink group (UCLA); IRG2‐6 is being conducted out of the Holden Lab primarily, substantially with investigators in IRG3; IRG2‐8 is being conducted jointly between the Holden Lab at UCSB and the IRG5 MSSR (HTS) facility with the Bradley Lab at UCLA; IRG2‐9 is being conducted within the Holden Lab and within the Stucky lab in IRG1. Current IRG 2 Research Projects:

IRG 2‐1: High throughput screening development in mammalian cells using macrophages and epithelial cells to develop paradigms for assessment of nanomaterials toxicity (André Nel, Saji George, and Tian Xia)

IRG 2‐2: Marine organismal nanotoxicology: Studying Nanomaterials’ (NMs) Interactions at the Molecular, Cellular, Organ, and Systemic Levels (Gary N. Cherr, Carol A. Vines, Elise Fairbairn, Brian Cole)

IRG 2‐3: Engineered nanoparticle biosorption, toxicity, and toxicity mechanisms in planktonic and biofilm bacteria (Patricia Holden, John Priester, Allison Horst, and Raja Vukanti)

IRG 2‐4: Dynamic energy budget modeling of toxic effects of CdSe quantum dots (Roger M Nisbet)

IRG 2‐5: Trophic transfer, bioaccumulation, and biomagnifications of engineered nanomaterials in basal levels of environmental food webs (Patricia Holden, John Priester, Rebecca Werlin, Randy Mielke, Galen Stucky, Gary Cherr, Jeff Zink, Angela Hwang)

IRG 2‐6: Electron microscopic methods for visualizing nanomaterials in biological specimens (Randy Mielke, Patricia Holden)

IRG 2‐7: Linking the physiochemical characteristics of carbon nanotubes to toxicological outcomes in vitro and in vivo (André Nel, Xiang Wang, Tian Xia)

IRG 2‐8: High throughput screening of lethal and sublethal effects of engineered nanoparticles to bacteria (Patricia Holden, Ken Bradley, Raja Vukanti, Bryan France, Angela Ivask, Robert Damoiseaux) ‐ Seed Project

IRG 2‐9: Evaluating the toxicity mechanisms of silver nanoparticles in bacterial systems with implications to wastewater treatment (Patricia Holden, John Priester, Aditi Singhal, Raja Vukanti, Allison Horst, Galen Stucky)

Integration: 1. IRG2‐1 with IRG5 (Bradley) for development and standardization of HTS screening protocols for

mammalian cells, zebra fish embryos and specific stress responses; with IRG1 (Zink), to acquire the next generation of nanomaterials within a combinatorial library and to use directed synthesis to test hypotheses related to dissolution and to electronic properties; with IRG6 to analyze the extensive data sets generated with HTS and to transfer data for use in the IRG6 (Cohen, Rallo, Liu) and risk ranking research.

2. IRG2‐2 with IRG2‐5, to complete joint manuscript publication; with IRG4 for development of NP

dispersion protocol in seawater and other aqueous media using various dispersants; with IRG1 (Zink


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group) who covalently linked a fluorescent tag (FITC) to ZnO which enabled visual tracing of the NP into embryos; with IRG5 (Bradley) where a trial HTS project with mussel hemocytes was conducted.

3. IRG2‐3 with IRG1 (Zink group, and Protocols Working Group with Godwin) where dispersion protocols

with bacterial growth media were developed then tested using HT‐DLS; with IRG3 (Cardinale) where protocols for sampling submerged biofilms from aquatic flumes were developed and used in aquatic mesocosm studies; with IRG1 (Mädler) where a manuscript regarding disagglomeration of TiO2 nanoparticles by bacteria was finalized and published; with IRG5 (Bradley) where aqueous culture conditions for bacteria are to be translated for use in HTS, and for coordinated recruiting of new postdoc in bacterial HTS research

4. IRG2‐4 with Holden (model development using existing data from IRG 2‐3) and with IRG3 (Cardinale) 5. IRG2‐5 with IRG1 (Stucky) and IRG2‐2 (Cherr) where a collaborative manuscript was finalized based

on this research, and with IRG1 (Zink) where additional studies with other NPs were begun.

6. IRG 2‐6 with IRG 2‐3 (Holden, aquatic biofilms), and IRG 3 (Cardinale, Lenihan) and IRG 4 (visualization using ESEM), where new methods in assessing NM aging in biological systems, mainly oriented around the use of energy dispersive spectral analysis of atom ratios, are being tested.

7. IRG 2‐7 with IRG 1 (Zink and Ji), to improve dispersion methods for multiwalled carbon nanotubes

including using bovine serum albumin (BSA) and assessing the role of hydrophobicity in regulating dispersion in cell culture media.

8. IRG2‐8 with IRG 5 (Bradley) where planning and trial of assays recruited and tested within IRG2‐3 was performed with HTS in a pilot project at UCLA’s MSSR for environmental bacterial screening.

9. IRG2‐9 with IRG1 (Stucky) and IRG5 (Godwin) where IRG1 methods for density gradient separation were recruited and modified for use with PHB‐producing bacteria, and where nano‐Ag was directly synthesized and characterized, then tested, for effects on monocultures and bacterial communities, and where variation in capping agents and use of surface enhance raman spectroscopy (SERS) were identified for use in a cross‐Center study of nano‐Ag versus ionic Ag effects on bacteria.

Major Accomplishments of IRG 2 (since March 2010): IRG 2‐1: Further advanced in vitro (cellular) and in vivo (zebrafish) toxicity screening, with cellular screening based on principles in the oxidative stress paradigm and zebrafish embryo screening based on population measures (survival, hatching, and physiological or morphological defects), to include assessments of Ag, Au, Pt, Al2O3, SiO2, ZnO and Q‐dot (CdSe/ZnS) nanoparticles. Expanded the assessment of TiO2 to address band gap effects (by tuning with Fe‐doping) and of crystal structure (e.g. anatase or rutile or mixtures), and screening entire TiO2 combinatorial library under light (photoactivated) and dark conditions in cell lines. Developed novel Electron Paramagnetic Resonance (EPR)‐based method for determining the specific reactive oxygen species generated during light activation of TiO2 particles in the combinatorial library. Acquired and characterized, with IRG1, a nano‐Ag combinatorial library; assessed toxicity in mammalian cells and in zebrafish embryos, plus acquired and screened a fish cell line, revealing differences across these three toxicity receptors. Overall, completed several combinatorial library studies including the above, as well as CdSe QDs, and the previously performed library with ZnO versus Fe‐doped (to control dissolution ) ZnO. With IRG6, advanced heat map as the display mode for the 5000+ data points per combinatorial library toxicity


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screening product, and used state of the art statistical approaches (i.e. z‐score) to define true effects within high throughput data sets. IRG 2‐2: Evaluation of toxicity of metal oxides (CeO2, TiO2, ZnO and Fe‐doped ZnO) to sea urchin embryos in seawater with and without various putative dispersants and organic matter (OM: including Bovine Serum Albumin or BSA, humic acid, alginate) where BSA and HA were retired based on non‐specific effects on embryo development, and low environmental relevance of BSA; thus alginate was selected for further study. Two of the NPs (CeO2 and TiO2) showed little toxicity but ZnO was toxic to developing embryos, causing developmental abnormalities at very low levels with dose‐ and exposure time‐, but not developmental stage‐, dependencies. FITC‐labeled ZnO entered cells and were shown to induce apoptosis in blastula‐stage embryos. Unlike studies in IRG 2‐1, Fe‐doped ZnO proved to be cytotoxic to sea urchin embryos in seawater, suggesting important environmental and organismal differences in the ability to mitigate ZnO toxicity by Fe‐doping. This is likely due to the aggregation in seawater, leading to more similar rates of dissolution in doped vs. non‐doped particles. Tested activity of metal oxide nanomaterials as substrates for multidrug efflux export proteins in fertilized sea urchin embryos, discovering that the metal oxide NMs tested are not substrates for the MDR transporters, nor do they perturb efflux activity. Explored the role of endocytosis in translocating NMs intracellularly during sea urchin egg fertilization, discovering that FITC‐labeled TiO2 was not endocytosed—a result that is still under investigation. Developed protocols for dispersing hemocytes of the marine mussel, Mytilus californianus, in preparation for HTS with these environmentally relevant cells. Developed and applied high content assays for assessing intracellular ROS, membrane damage, intracellular enzyme activity, and phagocytosis, discovering specific dose‐response relationships for ZnO, TiO2 and CeO2 and differential membrane disruption relationships (e.g. related to ROS for ZnO but not for CeO2). Discovered that phagocytotic activity was decreased with ZnO, but not with other MeO SRMs. Developed additional assays for future use in HTS including: superoxide production, and mitochondrial membrane potential (MMP). Initiated HTS trial at UCLA with mussel hemocytes and polystyrene beads as negative (non‐toxic) controls. Published a manuscript with IRG4 regarding dispersion of NPs in seawater and other environmental waters as a function of additives including alginate and other OM compounds; published a manuscript with IRG 2‐5, and submitted another manuscript. IRG 2‐3: Further evaluated, then augmented with iDNA data, optically‐based growth inhibition data for SRM metal oxides (CeO2, TiO2, ZnO; four bacterial strains, 2 gram positive, 2 gram negative) where it was reconfirmed that gram positive bacteria were found to be relatively more sensitive, and toxicity for all was enhanced in minimal, versus complex, aqueous media. Also, quantitatively assessed differential association of NPs with gram positive versus gram negative bacteria as shown by STEM‐EDS to develop a more detailed analysis of cell surface associations. Re‐assessed growth rates and further drafted a manuscript that is currently under revision prior to submission regarding differential effects of MeO SRMs on bacterial growth by strain, media and cellular association. Began studies of Fe‐doped versus undoped TiO2 effects on bacterial growth. Completed study of P. aeruginosa disagglomerating TiO2, using DLS, and quantitative high resolution microscopy; published related manuscript. Developed protocol for dispersing TiO2 in environmentally‐relevant bacterial growth media, worked collaboratively with IRG1 using HT‐DLS to expand protocol assessment. Applied protocols for culturing saturated (in flume mesocosms) and unsaturated bacterial biofilms with exposure to NMs; performed exposure studies with TiO2 (saturated biotilms) in collaboration with IRG3 (Cardinale), quantified biofilm cells/ macromolecules and associated TiO2, and collaboratively planned microbial community assays. As above, drafted one manuscript regarding planktonic bacterial growth studies, and published one manuscript regarding TiO2 disagglomeration by bacteria. Two other manuscripts are in final stages of preparation (unsaturated biofilm, and dispersion of TiO2 in bacterial growth media).


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IRG 2‐4: Developed dynamic energy budget model of effects of Cd(II) on bacterial growth, using growth curve data collected experimentally from IRG2‐3 for CdSe QD and Cd(II) growth inhibition studies published in 2009. Showed that assumptions involving reactive oxygen species (ROS) affecting controls improve the model fit to data. Completed calibration of model to new data generated by IRG2‐3 of optical density versus cell counts, using these results to calibrate data at high cell density regions of growth curves. Improved model using new assessments of lag times. Drafted and submitted manuscript for publication which is currently under revision for re‐submission for publication. IRG 2‐5: Completed all measurement interpretations related to simple trophic transfer study (bacteria to protozoans) of Cd(II) versus CdSe QDs where significant bioaccumulation in bacteria and biomagnification to protozoa are observed. Used combined STEM and growth modeling to reveal digestion impedance visualized in STEM images, and defined major discovery of this experiment as one of differential digestion due to feeding on QD‐laden bacteria. Researched related predator‐prey literature, submitted manuscript for publication, revised manuscript including acquiring additional data using high resolution TEM, and published revised manuscript which was a first showing biomagnification of engineered nanomaterials. IRG 2‐6: Completed acquisition of all STEM images of protozoa and bacteria associated with IRG2‐5 and completed EDS data acquisition. Began analysis of EDS data relative to controls with the objective of assessing atomic ratios of QD surface atoms (as resolved by EDS) at locations of QD cellular sequestration (e.g. cell membrane, mitochondria, food vacuoles, etc.). Ongoing is assessing efficacy of atomic ratio analysis for characterizing NP fate in biological tissues and cells. Completed STEM image acquisition for all bacteria‐NM conditions in IRG2‐4, with the objective of assessing cell damage by association of NMs with cell membranes. Acquired new STEM and TEM data with IRG2‐3 regarding detailed analysis of MeOs NMs with cell membranes; contributed these data to manuscript in preparation. Performed STEM and ESEM of bacterial biofilms (IRG2‐3 and IRG3, flume mesocosms) and algal mats for analysis of NM distributions in surface‐associated microorganisms; incorporated algal mat data into submitted manuscript with IRG3 (Cardinale lab). Started literature review of using EDS for aging analysis of NMs in tissues and organisms, in situ. Contributed to IRG2‐5 publication, as above. IRG 2‐7: Studied multiwalled carbon nanotube (MWNT, including as‐prepared, purified, and carboxyl‐modified) toxicity to human bronchial epithelial cells, finding no effects of metal impurities (in contrast to SWCNTs previously discussed) but rather that dispersion favored the production of key mechanism indicators, with the findings supporting prior reports in the literature of fibrogenesis by dispersed MWCNTs. Development of HTS assay to screen CNTs based on their physicochemical characteristics for causing cell proliferation, including refining dispersion protocols appropriate for conducting the assay. Performed fibrogenic study in mouse lung to establish effective doses of well‐dispersed MWCNTs which can therefore be applied for screening a wider range of MWCNTs with varying morphologies hypothesized to contribute to fibrogenesis outcomes. IRG 2‐8: Selected and recruited assays for evaluating sublethal effects of NMs on bacteria for translation to HTS. Defined criteria for performance, selected negative and positive controls. Completed assay recruitment for membrane potential, superoxide and dehydrogenase; membrane permeability and ROS were previously in‐hand. Planned full and pilot HTS campaigns; performed three HTS experiments in one campaign, showing the tremendous potential for HTS of bacteria for assessing potential environmental impacts of ecosystem relevance.


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IRG 2‐9: Studied effects of silver ions versus cysteine‐capped Ag NMs on gram negative bacterial growth and on sublethal indicators of toxicity. Challenged polyhydroxybutyrate (PHB)‐producing bacteria from activated sludge with Ag ions or cysteine‐capped Ag NMs during PHB synthesis phase, recruited density‐based separation method for isolating PHB‐rich bacterial populations, and quantified PHB production according to NM exposure and uptake. Reserved DNA for community analysis. Impacts on the Overall Goals of the Center: The overall goal of exploring mechanisms is most highly developed at this point for the mammalian cell lines in IRG2‐1 and also with the zebra fish in vivo research in IRG2‐1. The projects in IRG2‐1 have demonstrated the utility of the oxidative stress paradigm advanced by Nel et al. (2006), and the extraordinary value of a toxicity screening system that is based on knowledge of biochemical pathways and putative mechanisms of effects. Through IRG2‐1, the menu of NMs has widened beyond the initial MeO SRMs to now include gold, silver, carbonaceous, silica, copper, alumina, and semiconductor nanomaterials, with large datasets now feeding the modeling and risk ranking effort in IRG6. Also, the menu of conditions of exposure has widened to include light and dark, which is critical for a full assessment of photoactive TiO2 toxicity mechanisms (e.g. related to band gap modulation and thus ROS species and production). Similarly, the advancements in IRG2‐7 are showing how to work with carbon nanotubes in high throughput screening, the role of dispersion in MWCNT toxicity, and the relationship between indicators of fibrosis and fibrosis outcomes in animals. However, other projects are also demonstrating impact, including progress towards HTS‐oriented assays and screening results for dose‐response relationships in marine organisms (embryos and mussels) as well as for bacteria. The focus on mechanisms for these latter systems is progressing, having established effective doses and initial data sets regarding effects and nanomaterial associations with test organisms. Dispersion protocols and the use of environmentally‐relevant organisms and media, as methodological emphases, were advanced for all projects. Dispersant selection and performance was advanced with mammalian cells and embryos, where the latter, in seawater, effectively utilized alginate as a dispersant. Dispersion protocols and dispersants for bacterial culture media were tested with a final protocol developed around humic acid as a dispersant for TiO2. The substantive scientific findings are also enlarging, including beyond the theme of ZnO toxicity through dissolution and toxic effects of Zn ions, to showing the toxic effects of other metal‐shedding nanoparticles such as Ag and CdSe quantum dots. Doping ZnO with Fe was found to significantly reduce ZnO NP toxicity to mammalian cells, but not for sea urchin embryos which reiterates the value of many simultaneous detailed investigations using environmentally‐relevant organisms and media; the effects of doping with Fe on bacterial toxicity is to be tested. ZnO was found to enter and exert substantial toxicity to sea urchin embryos, and new methods are under development to assess cellular damage (e.g. oxidation and membrane damage). Bacterial toxicity research showed that ZnO and CeO2 are more growth inhibitory than TiO2; hypotheses for mechanisms and methods for testing differences between gram positive and negative strains are under evaluation, as well as for understanding the profound effect of oligotrophic verus rich media where toxicity was more enhanced in the former. A trophic transfer experiment (bacteria to protozoa) was completed, showing for the first time bioaccumulation and biomagnification of CdSe QDs in a simplified microbial food web, but, more importantly a differential effect of QDs on protozoan digestion of bacteria. The use of atomic ratios acquired with STEM‐EDS for tracing the integrity of CdSe QDs through trophic transfer was demonstrated. Two other high impact products include, for the first time, a DEB model applied to Cd(II)‐affected bacteria during growth, and also the discovery that bacteria can disagglomerate a common metal oxide which has implications for transport in porous media in the environment. Substantial progress was made within the marine and bacterial research areas to recruit, demonstrate and soon forward environmentally‐relevant assays into HTS. Contributions, including numerous talks and posters,


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were vigorously contributed from across IRG2 to the 2nd ICEIN conference and to other numerous national and international meetings. Major Planned Activities for the Next Year:

Continue to expand cytotoxicity studies in combinatorial nanomaterial library that includes Ag, Au, Pt, Al2O3, SiO2, ZnO and quantum dot (CdSe/ZnS) CeO2 , ZnO, CuO and CoO (with varying size), carbon nanotubes (with varying degrees of heavy metal contaminant and surface modification), and continue to analyze the data from in vitro and in vivo toxicity studies with combinatorial library nanoparticles including using computational expertise from IRG‐6 for (1) ranking the nanoparticles according to their toxicity (2) assessing the predictive power of in vitro studies, (3) start building up expert system required to generate structure activity relationships (in collaboration with IRG6). This study will help us in understanding toxicity paradigms other than oxidative stress paradigm and in generating the structure activity relationships.

Continue to study the cytotoxicity of iron doped TiO2 with cells under dark and illuminated conditions and understand the effect of iron doping on the abiotic and biotic reactive oxygen generation during light activation. Extend the evaluation of Fe‐doped TiO2 toxicity mechanisms related to ROS in light and dark conditions to bacterial cells.

Perform mice studies to assess in vivo effects of MWCNTs on fibrosis while measuring key biochemical indicators to support HTS assay development based on such indicators of observable adverse lung effects related to mesothelioma.

Evaluate oxidative damage and cell viability endpoints in sea urchin embryos, and in mussel hemocyte (immune and respiratory) cells exposed to a variety of nanomaterials including ZnO.

Expand NP range beyond initial triology of metal oxides for marine organisms / cells and bacteria.

Expand documentation of experimental protocols, with an initial emphasis on dispersant selection, testing, validation, and with the development of HTS testing approaches that are tuned to organisms / media selected for environmental research.

Use methods developed for HTS of nanomaterial toxicity to marine and bacterial cells; demonstrate and document performance and challenges.

Arrive at an initial explanation for differential growth inhibition of metal oxide NPs to gram positive, versus gram negative, bacteria, for across the SRM metal oxides and two media.

Advance NP aging characterization by TEM/EDS.

Prepare and submit several manuscripts for publication (at least three associated with IRG2‐1, one to two with IRG2‐2, two or more with IRG2‐3, one with IRG2‐4, one with IRG 2‐6, one or more with IRG2‐7, one from IRG2‐8, and one or more with IRG2‐9).

Expand project IRG 2‐5 to include Trojan horse NMs synthesized by IRG1, in order to control toxicity onset by pH‐triggered cargo release according to food vacuole acidity.


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IRG 3: Organismal and Community Ecotoxicology Faculty Investigators: Bradley Cardinale‐ UC Santa Barbara (now: U of Michigan)‐ Assoc. Professor, Freshwater Ecology Gary Cherr, UC Davis – Professor, Environmental Toxicology/Nutrition Jorge Gardea‐Torresdey, Univ. of Texas‐El Paso – Professor, Plant Toxicology Patricia Holden, UC Santa Barbara – Professor, Environmental Microbiology Hunter Lenihan, UC Santa Barbara – Professor, Applied Marine Ecology – IRG LEAD Edward McCauley, UC Santa Barbara – Professor, Freshwater ecology and Population Dynamics Roger Nisbet, UC Santa Barbara – Professor, Ecology and Population Dynamics Josh Schimel, UC Santa Barbara – Professor, Terrestrial Plant/Soil Ecology Number of Graduate Students: 5 Number of Undergraduate Students: 5 Number of Postdoctoral Scholars: 6 Goals of IRG 3: IRG 3 is organized to attain six major CEIN research goals, all of which focus on the development and testing of specific hypotheses regarding the impact of ENMs from the CEIN combinatorial library on organisms that are sentinels for key ecosystem services (primary production, human food production, biofiltration, deposit feeding, soil and material processing ‐ including ENM detoxification) and/or major ecological processes (species interactions that influence abundance and dynamics of sentinel organisms, trophic transfer, bioaccumulation, and biomangnification of ENMs; and food web dynamics that control the fate and transport of ENMs). It is not possible to attain our goals without integration within our IRG and with other IRGs. Our six major goals are to test hypotheses associated with: 1. CEIN Ecotoxicological Paradigm: The overarching goal of IRG 3 is to develop and test the hypothesis that the CEIN Ecotoxicological Paradigm is an efficient and powerful means of understanding the environmental implications of nanomaterials. The CEIN Paradigm is based on integrating information about ENM physiochemistry (IRG 1), potential physiological effects from QSARs (IRG 6), actual cytotoxic effects from HTS experiments (IRG 2/5), and fate and transport information (IRG 4) into a hierarchical framework that generates specific hypotheses about the biological impacts of ENMs on sentinel organisms, their populations, and communities.

The organisms we have chosen to study are sentinels for ecosystem services, or key ecological processes that control the provisioning of those services. Biological impacts of ENMs include alteration of demographic rates of individual organisms; population growth rates, abundance, and dynamics; coupled population dynamics and trophic transfer, bioaccumulation, and biomagnifications; and community composition, structure, and dynamics, including biodiversity material processing especially for ENMs. The results of our experiments used to test hypotheses about ENM toxicity are often used additionally to paramaterize Dynamic Energy Budget (DEB) models. DEB models are used to predict the impacts of ENMs at high levels of biological order (populations, communities, and ecosystem) when the only data available are from tests of whether and how ENMs modify energy utilization especially for growth and reproduction. Research conducted during this reporting period advanced development of the CEIN Ecotoxicological Paradigm through interactions between IRG 3 and IRG 1, 2, and 4; as well as work within IRG 3. For example, experiments using freshwater mesocosms, as well as a series of microcosms experiments in


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marine and terrestrial plan and soil media, that tested specific hypotheses about ZnO, TiO2, and CeO toxicity (described in detail below), were designed based on information provided by IRG 1 about metal oxide NM physiochemistry, and thus hypothetical bioavailability. These experiments also required information provided by IRG 4 about ENM fate and transport – specifically aggregation, dissolution, binding on soil and marine sediment – and thus actually bioavailability. Further advances in CEIN integration must come from greater integration of HTS and QSAR work focused on the sentinel organisms we use in IRG 3. Listed below are a number of projects that have been developed to develop our Paradigm. We have also devised a plan for better collaboration and integration across IRGs 1‐5.

2. Individual level effects of ENMs in microcosms: We are testing specific hypothesis concerning how sentinel organisms take up, bioaccumulate, and biotransform ENMs directly from the environment; testing whether ENMs cause genotoxicity or influence the demographic performance or vigor of individual sentinel (or other) organisms. Our hypotheses were generated when possible from QSAR results from the literature, and results from IRG 2 (physiology/histology) and IRG 4 (Fate and Transport). We hope to use HTS results in collaboration with IRG 5 in the future, but in the meantime have been developing slow‐through‐put protocols for our sentinel species that we hope to execute in HTS. When information and data necessary for generating hypotheses is not available from CEIN, we have looked to the literature for experimental work that is most useful, including QSAR work on ENMs and micro‐scale contaminants, for example ionic Zn and titania. Work in this period focused mainly on terrestrial plants, the sentinels of human food production; and marine amphipods, sentinels for sedimentary deposit‐feeding. Experiments included uptake and biotransformation experiments with ZnO, TiO2 and CeO2 in soybean, tomato, alfalfa, and corn; effects of ZnO and TiO2 on vigor (root and stem growth) of mesquite; biotransformation of ZnO and TiO2 by soil bacteria; and growth, respiration, filtration, and ENM bioaccumulation by marine mussels exposed to ZnO. ENM processing and biotransformation by marine mussels was also examined. Experiments with mussels provided a wealth of data for development and parameterization of mussel‐based DEB models. Substantial advances were made within IRG 3‐6 (a project recently transferred to IRG 2) to generate two novel systems for marine phytoplankton, one for marine mussels, and another for terrestrial soil bacteria. These two precursors to potential HTS protocols could in the future provide a foundation for generating specific hypotheses about effects of CEIN 2nd phase ENMs on these to key sentinel species. A complete description of work to be accomplished with the 2nd phase ENMs is provided in the March 2011 IRG 1‐4 Planning Document. However, future advances will be depend on greater integration and help from IRG 2 in providing information about ENM‐related cytotoxic mechanisms in sentinel organisms. Such mechanisms form the basis of the hypotheses we test in IRG 3 experiments. To date, we have relied on integration of efforts within IRG 3, depending heavily on the Prof. Cherr at UC Davis for developing methods that will lead to high content generation. 3. Population level effects of ENMs in microcosms: Our microcosm work is designed to test specific hypotheses regarding the effects of ENMs on population growth rates and dynamics of sentinel species. Primary production and respiration by whole populations is also measured in response to ENM exposure. Hypotheses are generated about the effects of ENMs on population growth rates based on results of HTS experiments, QSARs, and tests conducted at the level of the individual as described above. In this reporting period, a large number of tests were conducted to assess whether ZnO and TiO2 influenced population growth rates in marine phytoplankton, freshwater phytoplankton, soil bacteria, and marine amphipods. Hypotheses concerning the genotoxicity of ceria were also tested in terrestrial


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plants. Hypotheses used to test effects of ENMs on marine phytoplankton were generated in part from results of QSARs for E. Coli bacteria reported in the literature. Hypotheses tested in all environmental media‐ soil, freshwater, and seawater ‐ were also generated from information provided by IRG 1, 2, 4, and 5 about the dispersal potential and bioavailability of each ENM. We report the most important of these results in this summary report. 4. Community level effects of ENMs in microcosms: Work at the community level addresses hypotheses regarding the impact of ENMs on soil bacterial (Bacteria; bacteria‐protista) communities; freshwater benthic stream (biofilm) communities, and freshwater plankton (phytoplankton) communities; marine benthic cyanobacterial communities. 5. Effects of ENMs on trophic‐transfer and coupled populations in microcosms and mesocosms: Three focal community‐level effects tested in microcosms or mesocosms by IRG 3 involve trophic‐transfer of ENMs from one species to another (e.g., prey to predator); predator‐prey dynamics that measure the change in abundance of both predator and prey, as well as rates of predation (or grazing by herbivores); ENM and material (carbon and nutrients) processing by communities of organisms (e.g., multiple species of phytoplankton, bacteria, and biofilms) for the benefit or detriment of another organism; and finally competition between different organisms.

In this reporting period, our experiments in this context involved the transfer of TiO2 ENMs from freshwater algae to biofilms; competition between freshwater algae and bacteria; grazing on TiO2‐contaminated freshwater algae by stream snails; the trophic transfer of ZnO ENMs from phytoplankton to marine grazers (i.e., mussels in benthic systems; copepods in pelagic systems); the trophic transfer of TiO2 from marine phytoplankton to copepod grazers; the impact of ENMs in soil bacterial communities; and the development of a new coupled population dynamic mesocosm system, consisting of freshwater phytoplankton and Daphnia magna grazers in a sophisticated mesocosm system containing and exposing phytoplankton and grazers separately and together in inter‐connected microcosms. Like our work with marine mussels, the new freshwater mesocosm is designed in part to fully inform DEB modeling. We also designed and are developing a marine mesocosm that will examine trophic transfer and bioaccumulation in a simplified, coupled benthic sedimentary and pelagic food web consisting of phytoplankton, mussel filter‐feeders (rocky reef habitat), amphipod deposit feeders (sedimentary), and spiny lobster top‐predators. In collaboration with IRG 4, the marine mesocosm will examine fate and transport of a second phase CEIN ENMs (CNTs, CuO, AGm Pd, Pt, or QDOTs) to be determined from high content and microcosm work described above. 6. Dynamic Energy Budget Modeling: DEB models provide a quantitative framework for extrapolating the effects of ENMs on individuals, based on result of our HTS and microcosm studies, to population, community, and ecosystem‐level processes (population dynamics, species interactions, energy transfer). These models generate energy budgets associated with fecundity, egg production, growth, and mortality. The DEB models being developed so far focus on three of our sentinel organisms, marine mussels, freshwater phytoplankton, and Daphina. These models (1) generalize the effects of ZnO ENMs on mussel populations, mussel biofiltration, bioaccumulation of ENMs in marine food webs containing mussels, and marine ecosystem‐level processes, especially nutrient and carbon cycling influenced by biofiltration; and (2) explain how yet‐to‐be‐determined ENMs influence coupled predator‐prey dynamics among phytoplankton and Daphnia, which in turn affect the fate and transport of ENM and carbon/energy budgets associated with freshwater food webs.


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IRG 3‐1: Impacts of Engineered Nanomaterials on Marine Ecosystems (Hunter Lenihan)

IRG 3‐2: Toxicity and Uptake of Nanoparticles by Terrestrial Plant Species (Jorge Gardea‐Torresdey)

IRG 3‐3: Dynamic Energy Budget (DEB) Modeling to Support Design of Aquatic Microcosm and Mesocosm Experiments (Roger Nisbet)

IRG 3‐4: Nanotoxicology in Terrestrial Mesocosms (Patricia Holden, Joshua Schimel)

IRG 3‐5: The Impacts of TiO2 Nanoparticles on Freshwater Food Webs (Bradley Cardinale)

IRG 3‐6: Decoupling and Recoupling Plant‐Herbivore Systems to Determine the Fate and Impact of Nanomaterials in Freshwater Environments (Ed McCauley, Roger Nisbet)

Major Accomplishments of IRG 3 (since March 2010): Goal #1: Major accomplishments associated with developing the CEIN Ecotoxicological Paradigm were the development of conventional screening protocols for marine phytoplankton and mussel hemocytes by Prof. Gary Cherr and his group, and Lenihan‐Miller’s group, using seed funding. These systems will provide the basis for high content screening in the next reporting period of CEIN research. The phytoplankton and mussel HTS work will tests for specific cytotoxic responses (cell membrane integrity, cell viability, ROS, and mitochondrial function) to ceria, titania, ZnO, and all second phase CEIN ENMs, beginning with SWCNT, MWCNT, and carbon black. Additional accomplishments included a collaboration with IRG 1 to design and select second phase ENMs (excluding QDOTs, which are to be determined later) for HTS and microcosms experiments, as well as fate and transport experiments and modeling in IRG 4. In addition, we designed a new freshwater micro‐ and mesocosm system that allows testing of hypotheses about coupled population dynamics, as well as fate and transport, all for the purpose of parameterizing DEB models built specifically for the freshwater food web consisting of phytoplankton, Daphina, and bacteria. Finally, we developed an integrated system of analysis that couples marine mussel HTS experiments with individual demographic impacts and material/ENM processing through biofiltration, the data from which will be used in marine biolfiltration DEB modeling. Goal #2: Work on the individual level effects of ENMs in microcosms across freshwater, marine, and terrestrial plant systems found that in general ZnO ENMs are toxic to the sentinel organisms we examined, always reducing their growth, fecundity, and/or survival (Miller et al. 2010; Hanna et al, submitted; Hanna et al., in prep). In contrast, ceria usually had only moderate negative effects on individual performance (Hernandz‐Moreno et al. 2010a; de la Rosa et al. 2011). TiO2 had no negative effects in some tests, negative effects in some cases, or in the case of freshwater snails (sentinel grazers of freshwater algae), had positive effects on growth but no effects on survival and reproduction (Tsai‐Simek et al., in prep.). The underlying mechanism for these general results appears to be that ZnO releases Zn+ ions as ZnO ENMs dissolve in the media (seawater or soil) or are biotransformed in plants thus releasing Zn+ ions (Lopez‐Moreno et al. 2010b). Free Zn+ are often cytotoxic or reduce the uptake of other trace metals. Neither TiO2 or ceria appears to readily dissolve in the environment releasing ions, nor are they biotransformed in plants (Keller et al. 2010; de la Rosa et al. 2011). Work with marine amphipods, sentinels of sedimentary deposit‐feeding, indicated that ZnO ENMs are highly toxic, reducing survivorship drastically, but that marine sediments can greatly mitigate toxicity by binding the ENMs, or probably more accurately free Zn++ that dissolve from the ENMs, on attached organic molecules (Hanna et al., submitted). Toxicity to amphipods (LC50) of ZnO was observed in concentrations as low as 1.4 mg L‐1 in water but ZnO was only slightly toxic above 200 mg L‐1 in sediments. Experiments with mussels exposed to ZnO introduced in the mussel’s food (phytoplankton) found that this ENM reduced the individual performance of mussels, in terms of growth and fecundity,


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at even very low concentrations (0.5 mg L‐1 in water = 5 ug g‐1 of phytoplankton tissue) (Hanna et al., forthcoming). This is a direct form of trophic transfer that provides further evidence that uncoated ZnO ENMs have important negative environmental implications in most ecosystems.

Work in terrestrial plants, both sentinels for human food production and other species, also found that ZnO ENMs reduced root and stem growth, and caused wilting and chlorosis associated with production of various stress enzymes even at low concentrations of 500 mg L‐1 water in hydroponic systems (Lopez‐Moreno et al. 2010b). In some cases, ceria also generated the same results, and like ZnO, caused genotoxicity in a sentinel food plant, soybean (Lopez‐Moreno et al. 2010a). However, most often ceria as well as TiO2 were not toxic to plants, in large part we hypothesize because the plants store and do not biotransform the ENMs, thus not releasing toxic Me ions (Hernandez‐Viezces et al. 2011, de la Rosa et al. 2011). Goal #3: Experiments to test hypotheses associated with population level effects of ENMs in microcosms supported results of the individual level experiments in showing that ZnO ENMs reduce population growth rates in most sentinel species that we studied. Such was the case with four species of marine phytoplankton (Miller et al. 2010). Experiments examining population level effects of TiO2 showed that this ENM increased population growth rates in 18 freshwater phytoplankton species (Kulacki and Cardinale, submitted), but subsequent tests exhibited high species specificity in population responses, showing decreased population growth rates caused by increasing respiration rates in some species, but increasing growth rates in other species caused by enhancement of gross primary production (Cardinale et al., in prep). TiO2 exposure under even high concentrations in seawater (10 mg L‐1) did not influence marine phytoplankton growth rates, but as hypothesized, when exposed to UV‐light levels characteristic of surface waters on sunny days, even low concentrations of titania (< 0.2 mg L‐1) caused substantial reduction in population growth (Miller et al., in prep). UV exposure of titania is thought to generate ROS in many types of cells. However, specific toxic pathways will be explored in HTS under development. Goal #4: Experiments to test hypotheses regarding the community level effects of ENMs were conducted in soil microcosms with bacteria as the sentinel organisms. Results found that ZnO (at concentrations of 0.05, 0.1 and 0.5 mg g‐1 soil) but also TiO2 (0, 0.5, 1.0 and 2.0 mg g‐1 soil) reduced the diversity of soil bacteria (Ge et al. 2011) and also altered soil microhabitat structure with negative effects on bacterial abundance (Holden et al., in press). Experiments in freshwater mesocosms containing simplified food webs showed that while TiO2 did not affect algal population dynamics, algal community composition did significantly affect the fate of TiO2 in biofilms. Start‐up work for the new Freshwater IRG 3‐5 researcher Dr. Ed McCauley involved the designing of an integrated microcosm‐mesocosm system that will allow for rapid, simultaneous experiments at population (demographics and dynamics) and community (species interactions, nutrient cycling, and mioaccumulation/biomagnification) levels to quantify ecotoxicological effects of CEIN SRM MeOs, as well as on selected the second phase ENMs. Dr. McCauley’s work will allow for close integration of ecotoxicity testing and DEB (IGR 3‐3) modeling in CEIN. Goal #5: Experiments examining trophic‐transfer in coupled populations were conducted with TiO2 in freshwater and marine systems composed of phytoplankton and grazers. In freshwater, the hypothesis that increasing the concentrations of titania exposure to phytoplankton would increase tissue concentrations in Daphina (gazer) was supported (Kulacki and Cardinale, submitted). In the marine system, we hypothesized that copepod (grazers) tissue levels of tintania would increase with the concentration of titania in phytoplankton cells, and that when exposed to relatively high UV light,


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individual growth rates and fecundity of copepods would decrease. Both hypotheses were supported by results of the experiment (Miller and Lenihan, in prep). We developed a collaboration with a copepod ecologist and toxicologist, Prof. G. Bielmyer of Valdosta State University, who will conduct the remainder of our coupled phytoplankton‐copepod experiments designed to test similar hypotheses will all CEIN second phase ENMs. Goal #6: We used DEB models as sophisticated regression tools to estimate no‐effect‐concentrations in marine phytoplankton populations exposed to ZnO ENMs (Miller et al. 2010). We also developed a mussel DEB model that we are parameterizing with several data streams generated from marine microcosm experiments. DEB model development and parameterization was also used in the design of the new freshwater mesocosm experiments being erected by Prof. McCauley in coordination with Prof. Nisbet, as well as IRG 1 and 4. Impacts on the Overall Goals of the Center: IRG 3 is answering important questions with regards to the toxicity of ENMs in various environmental media at levels of the individual to communities. We are addressing the key questions with regards to uptake of bioavailable ENMs; their processing and biotransformation; trophic transfer; and the results of their toxicity in terms of the demographic performance of sentinel marine, freshwater, and terrestrial organisms; whether and how they influence species interactions; and how they affect community composition, structure, and dynamics. We are testing many novel ecotoxicological questions but our main goal is to develop and test the CEIN Ecotoxicological Paradigm. So far, we can state that: (1) ZnO is under most conditions toxic to most sentinel species, mostly due to the Zn2+ ion exposure rather than the ENMs themselves; (2) that TiO2 is rarely toxic unless under high UV light conditions when is becomes photo‐toxic, or when it modifies communities of soil bacteria; and that ceria is rarely toxic but does cause genotoxicity in plants; (3) the presence of organic matter in marine sediments greatly decreases the bioavailability and toxicity of MeOs in marine ecosystems; (4) that uptake and biotransformation by plants and biofiltration by marine suspension feeders substantially modifies or sequesters MeO ENMs making them less bioavailable to most species but often leads to bioaccumulation in some grazers and predators; and that community biodiversity can influence the bioavailability of ENMs within food webs contained within mesocosms. Our work in this IRG is leading to the development of powerful DEB models that will generalize the ecological impacts of ENMs. Major Planned Activities for the Next Year: Critical next steps necessary for CEIN to attain our first goal of developing the CEIN Ecotox Paradigm is to focus HTS experiments in IRG2/5 on the sentinel organisms used in IRG 3 experiments. This will allow IRG 3 to generate more specific hypotheses about the effects of whole individuals based on cytotoxic mechanisms associated with CEIN’s first phase MeOs (ZnO, titania, and ceria) already examined; and the second phase CEIN ENMs (CNTs, CuO, as well as variants of Ag, Pd, Pt, and QDots). In this reporting period, we generated and tested ecologically relevant hypotheses based on mechanisms of ENM toxicity described in the literature, of which there are relatively few, but more generally information on the macromolecular and ionic phase toxicity of the first phase MeOs (see appendix) mainly ZnO, tintania. As described in our soon to be released March 2011 Planning Document, we will focus on marine phytoplankton screening of all CEIN 2nd phase ENMs followed by population growth rates of marine phytoplankton exposed to all 2nd phase ENMs. We will conduct whole mussel rapid screening of all second phase ENMs to test whether they influence respiration and biofiltration. Trophic‐transfer studies will be conducted with marine phytoplankton and copepods and we will conduct amphipod bioassays with ENMs recommended by IRG 1 and 4. Mesocosms experiments will be developed for freshwater and marine food webs, and soil‐bacteria systems. Finally, DEB models will be generated for marine mussels.


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IRG 4: Fate & Transport of Nanoparticles Faculty Investigators: Yoram Cohen, UCLA – Professor, Chemical Engineering Arturo Keller, UC Santa Barbara – Professor, Environmental Biogeochemistry – AREA LEAD Hunter Lenihan, UC Santa Barbara – Associate Professor, Marine Biology Ponisseril Somasundaran, Columbia University – Professor, Materials Science Sharon Walker, UC Riverside – Assistant Professor, Chemical and Environmental Engineering Number of Graduate Students: 6 Number of Undergraduate Students: 1 Number of Postdoctoral Researchers: 3 Goals of IRG 4: The main goal of IRG 4 is to generate the experimental data that will allow the prediction of the concentration of nanoparticles (NPs) at which organisms will be exposed to in different environmental compartments. This includes determining the mobility of the NPs, their reactivity, bioavailability, and persistence under various environmental conditions, including those in CEIN mesocosm studies. Organization and Integration of IRG 4 Projects: Current IRG 4 Research Projects:

IRG 4‐1: Photoactivity of Nanomaterials in Natural Waters (Arturo Keller)

IRG 4‐2: Role of Material Properties and Environmental Condition on Nanoparticle Aggregation (Arturo Keller)

IRG 4‐3: Effect of Wettability on the Transport and Fate of Metal Oxide Nanoparticles (P. Somasundaran)

IRG 4‐4: Packed Bed Column, Parallel Plate Flow Cell, and Radial Stagnation Point Flow Chamber Transport Studies Using SRMs (Sharon Walker)

IRG 4‐5: Attachment of Nanoparticles to Natural Surfaces Under Different Aqeuous Solution Chemistries (Arturo Keller)

IRG 4‐7: Interactions Between Biological Surface and Engineered Nanoparticles (Arturo Keller, Milka Montes)

Broadly, the IRG is divided into projects that determine the (1) mobility of CEIN NPs in different media; (2) their reactivity and persistence; and (3) their bioavailability. Since NPs are essentially colloids, the mobility studies are based on colloidal science, although NPs have been shown to be a distinctly different subset due to their enhanced physicochemical characteristics. IRG 4‐2 studies the factors that control aggregation in open waters (fresh and sea water); IRG 4‐5 determines the attachment of NPs onto natural surfaces such as sands and clays in marine and freshwater systems such as those used by IRG 3; and IRG 4‐4 determines the movement of NPs in porous media such as soils, along with groundwater. With regards to their reactivity, IRG 4‐1 is focused on the photoactivity of different NPs under environmentally relevant concentrations and conditions. IRG 4‐3 evaluates the surface energy of various NPs, which can be used to understand their interactions with other NPs, minerals and biomolecules. As a


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service to the CEIN, IRG 4‐2 also determines the rates of dissolution of NPs in IRG 3 media, since this is an important fate that can determine their persistence. In 2010 we began a new project, IRG 4‐7, which is focused on the biological uptake of NPs in different media, interacting closely with IRG 3. This will provide information on the actual bioavailability of various NPs. NPs are being tracked as they attached to phytoplankton, which then are taken up by mussels and distributed to various soft and hard tissues within the mussels. Other biological systems will be studied in the future. In early February 2011, we (IRG 4 in collaboration with other UCSB CEIN researchers) hosted a 1‐day workshop to determine how best to incorporate Life Cycle Assessment (LCA) into CEIN research. LCA determines the potential emissions of NPs into different environmental media at various stages within the life of a nanomaterial. The workshop included 5 leading researchers working on LCAs. At the end of the workshop, we identified the research gaps needed within LCA for nanomaterials and in the connection of LCA with fate and transport. LCA also needs to consider the broader environmental implications of nanomaterials. A PhD student working on these questions at UCSB has been identified, and we will be seeking seed funding from the UC CEIN to support her work as part of a new project within IRG 4. In addition, a Working Group on Nano Fate and Transport has recently been formed by Cohen and Keller, which includes researchers from IRGs 4 and 6. The working group will be developing collaboratively the various elements of the multimedia fate and transport model, incorporating the experimental data generated by IRG 4. As needed, the Working Group will bring in or consult with other IRGs with regards to issues that involve connecting the exposure model (i.e. the concentration at which NPs are likely to occur in different media) with the effects models (toxicity and other impacts). The Working Group will also determine how best to connect LCA with Fate and Transport. Major Accomplishments of IRG 4 (since March 2010): In 2010 a major study (Keller et al., 2010) was conducted by IRG 4‐2 that evaluated the state of metal oxide (MeO) NP aggregation and the rate of aggregation and sedimentation in nine natural waters, to inform the entry points of MeO NPs into the environment from various uses within their life cycle. For example, we determined that TiO2 NPs that may be washed off surface coatings by stormwater, or CeO2 NPs carried by runoff, will form relatively stable suspensions due to the high NOM and ionic strength (IS) of stormwater. Similarly, MeOs that are washed off from sunscreens and pass through wastewater are also likely to be suspended, but will begin to aggregate and sediment out sooner due to the higher IS, despite the high NOM in wastewater. If the stormwater or wastewater is discharged to a freshwater body (e.g. rivers, lakes), the MeO NPs are again likely to remain suspended for tens of hours due to the low IS and significant NOM. However, in groundwater, the high [Ca2+] and lower NOM leads to high rates of aggregation (minutes to hours) and sedimentation. In seawater, the high IS comes from [Na+], but leads to similar high rates of aggregation and sedimentation. The theoretical basis for these observations was evaluated (Thio, Zhou and Keller, 2011) by carefully controlling water chemistry in synthetic aqueous matrices, and can now be explained by the specific role of each major cation and [NOM]. This knowledge will be used to calibrate and validate the NP aggregation model developed by IRG 6, which will form part of the CEIN multimedia model that serves to predict NP concentrations in various media. It also served to inform IRG 2 & 3 researchers as to the state of their MeO NPs during their toxicity studies. IRG 4‐2 also provided a protocol for maintaining the NPs suspended using alginate, so that IRG 2 & 3 studies could evaluate the difference between a stable suspension and a destabilized one (Fairbairn et al., 2011).


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Given that the toxicity of ZnO NPs is most likely due to the release of Zn2+, IRG 4‐2 conducted dissolution experiments in IRG 3 waters, including fresh and sea water. These studies showed that the ZnO NP dissolve within 12 hr, and thus helped to confirm that the toxicity was most likely due to Zn2+(Miller et al., 2010; Fairbairn et al., 2011). The dissolution rate was compared to that of bulk ZnO, which helped to confirm that the rate was considerably higher for the high surface area NPs. A study was also conducted to determine the rate of dissolution of Fe‐doped ZnO. Clearly, doping ZnO with Fe resulted in slower dissolution, but the Fe‐doped ZnO NPs also aggregated much faster in seawater compared to undoped ZnO (Fairbairn et al., 2011). Thus, the decreased dissolution rate is a function of two effects, namely doping and reduced surface area due to the higher state of aggregation. These results helped to inform IRG 2 and 3 studies using ZnO, and will also serve to parameterize the IRG 6 multimedia model. In the last few months IRG 4‐2 has been addressing two important issues for the CEIN. First, work is proceeding on evaluating the rate of aggregation of the new CEIN NPs (CNTs, Ag, Pd, Pt), recently received from IRG 1. Work to date indicates that these NPs tend to be stable in freshwater, except Pd NPs. They aggregate rapidly in seawater; work is underway to determine if alginate will stabilize them as it did for MeOs. As shown earlier by Wang, Keller et al. (2008), CNTs can be dispersed using NOM (in the form of humic acid). However, the dispersability of CNTs is a function of purity and additional research is being conducted to better understand the controlling factors. These new NPs will be studied in natural waters (including IRG 2 & 3 conditions) as well as controlled synthetic aqueous matrices to understand the controlling factors and be able to provide IRG 6 with the information needed for the multimedia model. The second issue is the role of NP properties, such as size, crystalline structure and morphology. Using IRG 1 TiO2 NPs, a combinatorial matrix of 16 different particle characteristics is being studied to determine the relationship between particle properties and aggregation. This follows the work done in 2010 where we showed conclusively that ZnO morphology was very important for understanding aggregation (Zhou and Keller, 2010). Spherical particles are much more stable than plate‐like NPs, probably due to the many contact points along the plane. This information must be considered in IRG 6’s aggregation model, to more accurately predict rates of aggregation and thus concentrations in different media. Project IRG 4‐5 studies the attachment of NPs onto mineral surfaces, using Atomic Force Microscopy (AFM) and Quartz Cell Microbalance (QCM) to measure the interaction forces. In Thio et al. (2010), we presented a novel approach for using AFM to measure these nanoscale attachment forces. Knowing these forces allows us to predict whether the NPs will be stably removed from the water column, or may be resuspended due to turbulence. This study was followed by another one (Thio, Zhou and Keller, 2011) in which we reported that the presence of NOM significantly reduces the attachment of the NPs to mineral surfaces such as sands (silica) and clays (mica), making the NPs and their aggregates more mobile. Thus, under most conditions the aggregated NPs will not deposit firmly on the sediments and are more bioavailable, both to sediment (benthic) organisms and those in the water column (pelagic). This information is important for the design of IRG 3 mesocosm studies and for the IRG 6 multimedia modeling effort. In the current phase of the project, the new NPs (CNTs, Ag, Pd, Pt) are being evaluated with regards to their attachment to mineral surfaces under natural conditions (NOM, IS, pH) using IRG 3 waters. In addition to AFM and QCM, we are exploring the use of Thin Layer Chromatography, since it may prove to be a much faster method for determining the deposition rates under various conditions. Project 4‐4 studies the transport of NPs through porous media such as groundwater in soils. The Walker lab completed a study on role of solution chemistry, nanoparticle concentration and hydrodynamic effects on transport of TiO2 NPs through porous media (Chowdury et al, 2011a,b in review). Ionic


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strength and pH were varied systematically, demonstrating a significant impact on the transport due to their role in the aggregation of the NPs and interaction with quartz sand. NP concentration also played a significant role specifically under chemically favorable conditions, leading to increased breakthrough of NPs due to blocking and subsequent particle‐particle repulsion. Increased flowrate resulted in greater elution of NPs due to hydrodynamic forces acting on aggregates and contributing to blocking. The key finding was the interplay between mechanisms that result in filtration, straining and blocking under different conditions. Quantitative results from this study for the various mechanisms will provide the information needed by IRG 6’s multimedia model. In addition, IRG 4‐4 contributed significantly to the development of an NP dispersion protocol for the CEIN (Chowdury, Hong and Walker, 2010), providing information on the conditions that are needed for a stable stock suspension. Current work in IRG 4‐4 is investigating the transport of TiO2 nanoparticles in more complex conditions, particularly, the presence of NOM and bacteria both of which are ubiquitous in porous media. This work also will serve to inform the design of terrestrial mesocosm studies in IRG 3 and the work by Gardea‐Torresday’s group in IRG 2 with terrestrial plants. IRG 4‐1 completed a comprehensive study of the photoactivity of four MeO NPs (TiO2, ZnO, CeO2 and Fe2O3) in a high throughput screening mode (Bennett and Keller, 2010). Some key findings include the order of photoactivity of these four MeOs (TiO2 > γ‐Fe2O3 > ZnO > CeO2) in DI water. Interestingly, the order is different in seawater (ZnO > TiO2 > γ‐Fe2O3 > CeO2). There is a significant decrease in photoactivity in seawater for most MeO NPs, due to their rapid aggregation which reduces surface area. CeO2 is essentially not photoactive and may actually quench photoactivity. Photoactivity may have several effects: it may produce reactive species that interact with the organisms, it can accelerate the rate of mineralization of nutrients, and it can change the rate of transformation of other water quality constituents, including pollutants. As was observed in NP aggregation and deposition studies, NOM plays an important role in controlling the photoactivity of NPs. IRG 4‐1 is studying the effect of NOM as well as the new CEIN NPs. In addition, Bennett in IRG 4‐1 discovered a new phenomenon which has not previously been reported in the literature, namely the photoinduced disaggregation of NPs (Bennett, Zhou and Keller, 2011 in review). This may have very significant implications for toxicology, since aggregates shed primary nanoparticles when irradiated with sunlight, and our preliminary experiments show that the released NPs can migrate through porcine dermal tissue. We have observed the photoinduced disaggregation for MeOs and CNTs. As with other IRG 4 projects, IRG 4‐4 is collaborating closely with researchers in IRG 3 to understand the effects of photoactivity in freshwater and seawater systems with different organisms; joint publications are under way. The work will also inform IRG 6’s multimedia model with regards to the effect of photoactivity in different media. Project 4‐3 focuses on the wettability (surface energy) of NPs, and their interactions with clay minerals and some organic molecules. Researchers in IRG 4‐3 have developed a novel non‐destructive technique to measure the surface energy of an as received solid surface down to the nano scale. For the first time, the surface energy values of different kinds of solids can be evaluated by a single and simple technique under ambient conditions. It is their hypothesis that the NP surface energy can be correlated to their interactions with other substances such as proteins and minerals, to their aggregation kinetics, and the resultant transport performance. They find that NPs that have very low surface energy aggregate fast, while particles with high surface energy dissolve and disperse well in water (Fang et al., 2010a). High surface energy leads to more toxicity for the bacterium N. Europaea (Fang et al., 2010b), relative to low surface energy. The next phase will look at the wettability of NPs from Madler’s group in IRG 1, and the interactions of MeO NPs with different clay minerals. This project has yet to fit in well with the needs of the UC CEIN.


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The seed project IRG 4‐6 that evaluated the removal of MeO NPs from mesocosm wastewaters was completed. The key finding was that by destabilizing the wastewater by shifting pH and/or increasing IS, large aggregates would be formed, which can either be removed by decantation or traditional filtration (Surawanvijit, Kim and Cohen, 2010). This information is being used to process mesocosm wastewater in IRG 3, and served as the basis for the aggregation model developed by IRG 6. A new project (IRG 4‐7) was started in June 2010, to explicitly address questions regarding the bioavailability of NPs, and in particular uptake and excretion. Dr. Milka Montes, who recently graduated from Dr. Gardea‐Torresdey’s group at UTEP, is now working with Dr. Keller’s group. She is working closely with Lenihan group (IRG 3) to measure the uptake of MeO NPs by marine phytoplankton (live and dead) and then the uptake of the phytoplankton by mussels. The mussels then distribute the MeO to various soft and hard tissues. Dr. Montes is determining the concentration and mass of MeOs in different tissues. The most important finding to date is the accumulation of significant amounts of MeOs in pseudofeces, which are excreted by the mussels. These pseudofeces with NPs are then available for other organisms to uptake, which can result in additional exposure to MeO NPs. After completing the work with MeOs, IRG 4‐7 will begin studying the uptake, bioaccumulation and excretion of the new CEIN NPs. This work will be extremely valuable for IRG 3, and will also inform the multimedia model in IRG 6. Impacts on the Overall Goals of the Center: IRG 4 is answering important questions with regards to the state of the NPs in various environmental media, and the rate of change between states. We are addressing the key questions with regards to the mobility, persistence, bioavailability and reactivity of NPs in actual environmental media. We are working with the NPs provided by IRG 1 in the first year (metal oxides), in the media used by IRGs 2 and 3 in their studies. We have also evaluated the behavior of the NPs in other relevant environmental media. Thus, we can state that : (1) ZnO is likely to dissolve rapidly (within days) in most natural conditions, so that the risk is mostly due to the Zn2+ ion rather than the NPs themselves; (2) MeO NPs aggregate rapidly in seawater and other high ionic strength media, but are much more stable in freshwater and other media high in NOM and low ionic strength; (3) the presence of NOM plays a major role in preventing attachment of the MeO NPs to mineral surfaces; (4) filtration of the NPs in groundwater and sediments may occur if they aggregate significantly, but mostly due to straining as opposed to attachment to the porous media; (5) the mobility of the MeO NPs is generally greatly increased by the presence of NOM, whether in seawater, freshwater or groundwater; (6) the photoactivity of the MeO NPs is a strong function of surface chemistry; (7) the wettability of the MeO NPs controls their rate of aggregation and their interactions with organic molecules; and (8) NP morphology plays an important role in controlling aggregation, with results so far indicating that a flat, plate‐like morphology aggregates much faster than a spherical particle. These results will be key for designing IRG 2 and 3 experiments, as well as for the modeling efforts in IRG 6. Major Planned Activities for the Next Year: The most important major activity will be the incorporation of a broader set of NPs from IRG 1’s library, particularly the new particles (CNTs, Ag, Pd, and Pt), as well as NPs with different sizes and morphologies. We also have launched the Nano Fate & Transport Working Group, which will facilitate the transfer of information from IRG 4 to IRG 6, improving the development of the multimedia model. We also expect to obtain seed funding for the proposed new project that will construct a Life Cycle Assessment framework for nanomaterials, providing much needed information on the emissions of NPs. This will improve our estimates of NP concentrations in different environmental media.


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IRG 5: High‐Throughput Screening For Biointerfacial Properties of Nanomaterials Faculty Investigators: Kenneth Bradley, UCLA – Assistant Professor, Microbiology – AREA LEAD Hilary Godwin, UCLA – Professor, Environmental Health Sciences Patricia Holden, UC Santa Barbara – Professor, Environmental Microbiology Shuo Lin, UCLA – Professor, Molecular, Cellular, and Developmental Biology André Nel, UCLA – Professor, Medicine; Chief, Division of NanoMedicine Donatello Telesca, UCLA – Assistant Professor, Biostatistics Jeffrey Zink, UCLA – Professor, Chemistry and Biochemistry Number of Graduate Students: 2 Number of Postdoctoral Researchers: 8 Goals of IRG 5: The overarching goal of IRG5 is to rapidly determine the specific features of NMs that govern their biological interfacial properties. To accomplish this, we are leveraging existing high throughput screening (HTS) capabilities at the UCLA Molecular Screening Shared Resource (MSSR). Several key cell types (e.g., animal, yeast, bacteria) for their toxicity outcomes using a variety of assays that probe direct cytotoxicity, as well as sub‐lethal induction of stress. Further, we are employing genomics‐based HTS in E. coli and S. cerevisiae in order to determine mechanisms of toxicity induced by NMs. Organization and Integration of IRG 5 Projects: Bradley and Damoiseaux provide general HTS expertise and work closely with project leaders and research teams to facilitate HTS experiments. Currently five active projects are being undertaken by IRG5 (5‐2, 5‐3, 5‐5, 5‐6, 5‐8, and 5‐9). These projects address NM interactions with mammalian, yeast, and bacterial cells. In addition, there is scientific integration between IRGs 1 and 5 in nanomaterial characterization in different media as well as in projects IRG1‐6 and IRG1‐8 and strong collaboration on data analysis with IRG6. Collaborations also to bridge IRG 2 with several HTS projects addressing mechanisms of toxicity in mammalian (2‐1 and 2‐7), marine (2‐2) and bacterial systems (2‐8). Assays that measure the response to nanomaterials in mammalian cells are performed in collaboration with Nel (IRG 5‐5, 5‐6) and Bradley (IRG 5‐2). Godwin, oversees yeast (IRG5‐3) and bacterial (5‐8) assays in collaboration with Bradley. Finally, new HTS assays are being developed based on cells from marine organisms (mussel hemocytes, Cherr), and future plans involve development of HTS assays for marine organisms (Lenihan and Godwin). Core Function:

IRG 5‐2: Validation of Cell‐based Assays for High Throughput Screening to Determine Toxicity of Nanomaterials (Bryan France, Robert Damoiseaux, Ken Bradley)


IRG 5‐3: High‐throughput Characterization of Toxicity and Uptake Mechanisms of CEIN Nanomaterials in S. cerevisiae (Elizabeth Suarez, Robert Damoiseaux, Ken Bradley, Hilary Godwin) ‐ planned to end Spring 2011.

IRG5‐5: Understanding of Property‐activity Relationships of Different Silica Phases and Crystalline Polymorphs in the Toxicity of these Materials. (Haiyuan Zhang, Tian Xia, André Nel)


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IRG 5‐6: Development of High Content Screening of Nanoparticle Toxicity using in vivo Zebrafish models (Yan Zhao, David Schoenfeld, Saji George, Tian Xia, Sijie Lin, André Nel)

IRG 5‐8: High throughput bacterial screening for the characterization of toxicity of nanosized particles and materials (Angela Ivask, Robert Damoiseaux, Kenneth Bradley, Hilary Godwin)

Major Accomplishments of IRG 5 (since March 2010): IRG5‐2: 1. Lung epithelial cells (BEAS‐2B) were transduced with the luciferase reporter pathways and

functionality verified with the positive stimulants (refer to fields below). These cell lines have been scaled up to HTS format for screening with the eight SRMs (metal; gold, silver, platinum & metal oxides; aluminum, zinc, silica, iron & CdSe/ZnS quantum dot). Preliminary results show ZnO downregulating CRE, HIF, NFkB, SMAD, SRE, and AP1 transcription factors in a time and dose dependant manner which perhaps indicates sublethal toxicity and dysfunction in cAMP and inflammatory regulation. Also, the quantum dot surprisingly shows a time and dose dependant induction of the MAPK pathway. This data set has been communicated to IRG6 for statistical significance and modeling and resulted in an EST publication.

2. An alternative assay read out was tested using 10mM luciferin substrate in the culture media in transduced RAW 264.7 reporters known to respond to zinc oxide. Initial experiment did not yield a responsive readout.

IRG5‐3: 1. A full‐library HTS was performed using the S. cerevisae genome‐wide knockout collection to identify

genes that influence response to positively charged nanoparticles. The screen was performed in quadruplicate using 62 nm amino coupled polystyrene (PS‐NH2) as the challenge agent. This procedure yielded 268 robust knockout mutants, where the presence of the gene is involved in the cell’s response to the nanoparticle. Secondary assays were run on these hits to determine dose response from 0‐100 ug/mL 62 nm PS‐NH2. The IC50 valuels for each mutant was calculated and compared to the WT‐like strains. The reconfirmation rate was 64% for PS‐NH2‐resistant strains, with 155 out of 242 showing altered IC50 values. The initial reconfirmation rate for the sensitive strains was 23.5% (52 out of 221). The sensitive reconfirmation results seemed low, prompting an examination of the IC50 sensitive results which yielded identification of 53 additional strains with > 10 ug/mL differences in IC50s from the WT‐like strains, but with very wide 95% confidence intervals overlapping the WT‐like 95% confidence intervals due to steeep dose‐response curves.

2. Reconfirmed mutants were analyzed by gene ontology (GO) functional analysis and the PANTHER (www.panther.org) protein classification tool. Most of the sensitive mutants encoded genes knocked out that were involved in metabolism pathways of proteins and nucleic acids. One of the knockouts is of a serine/threonine kinase which is identified as being involved in ten identified signaling pathways involved in responses to stimuli or cell morphogenesis. Most of the robust mutants had genes knocked out that were involved in the metabolic processes for proteins and reactive oxygen species, mitosis, and signal transduction. One of the knockouts had the RAS1 gene deleted, which is involved in six signaling pathways. Preliminary gene interaction plots show the reconfirmed genes have significant numbers of interactions within either the sensitive and robust genes and between each other.

3. To determine if the reconfirmed yeast knockouts were homologous to genes in organisms of interest (E. coli, zebrafish, mammals/mice), a homology search was conducted using the reconfirmed genes and the web search tools OrthoMCL(www.orthomcl.org) and Homologene from the NCBI http://www.ncbi.nlm.nih.gov/homologene). For S. cerevisiae to E. coli homology, 15 genes from the sensitive mutants and 13 genes from the robust mutants showed homology. The sensitive


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mutant genes were mostly RNA processing, several transporters, and the TGF beta pathway involved in cell defense. The robust mutant genes were involved in uptake and metabolism of sugars and detoxification of metabolism byproducts. For S. cerevisiae to Danio rerio (zebrafish) homology, 17 genes from the sensitive mutants and 7 genes from the robust mutants were found. The robust genes in zebra fish were involved in protein metabolism, endocytosis, and sensory perception. The sensitive genes in zebrafish were involved in protein metabolism, purine biosynthesis, cation transport, and the wnt signaling pathway. For S. cerevisiae to mouse (Mus muscululs) homology, 17 homologous genes representing genes knocked out in robust strains were found. Most of the genes were involved in protein metabolism, mitosis, and signal transduction. Of these, four were found distributed among five signaling pathways. For genes knocked out in sensitive strains, 9 genes homologous to mice were found. Of these, most were in protein and nucleic acid metabolism.

4. To compare the toxicity of 62 nm PS‐NH2 to other substances, dose response tests were conducted with bleomycin and polymyxin B. Bleomycin is a cancer drug that disrupts DNA and the cell membrane and has known toxicity to yeast. Polymyxin B is a bacterial toxin that disrupts the cell membrane and has a known toxicity to yeast. The IC50 of bleomycin in our optimized media is 15.5 nanograms/mL. Polymyxin B IC50 in our media is 10 ug/mL. Per literature values, cisplatin, another cancer drug, has an IC50 of 51 ug/mL in YPD against the same WT strain of yeast. In comparison the IC50 of 62 nm PS‐NH2 is 38 ug/mL in rich YPDA at pH 7, suggesting these nanoparticles are very toxic.

5. Toxicity of 25 nm titanium nanoparticles from the SRM library was determined on WT yeast. The IC50 of the titanium was 543 ug/mL with 1% FBS in a 10 % water vehicle indicating that it may be used for future genome‐wide screens under these conditions. Specifically, it was found that addition of FBS was required for dispersion.

IRG5‐5: 1. We established a provisional silica nanoparticle library that includes amorphous colloidal silica,

amorphous fumed silica, mesoporous silica, silicalite and quartz. Their respective amorphous or crystalline phases, primary size and hydrodynamic size were confirmed by XRD, TEM and DLS. This activity is described in IRG 1‐6.

2. The MTS, LDH and ATP single assay screening results indicated an increased hazard potential for amorphous fumed silica and quartz compared to amorphous colloidal silica, mesoporous and silicalite that showed little or no toxicity. Multi‐parametric HTS screening also confirmed that fumed silica and quartz induced effects on plasma membrane leakage, increased intracellular calcium influx, and the generation of hydrogen peroxide and superperoxide at 1‐6 and 24 HR. Besides, fumed silica also led to mitochondrial membrane depolarization at 24 h. In contrast, amorphous colloidal silica, mesoporous silica and silicalite did not show any toxicological effects in the HTS. This research is now being persued from the perspetive of the silanol and siloxane groups that determines also surface reactivity of silica nanoparticles. Fumed silica is a high volume produced material that is used for several industrial applications.

IRG 5‐6: 1. We have successfully screened the compositional transition metal NP library and demonstrated that

CuO, Ni, NiO and SWCNTs with a relatively high Ni content can inhibit zebrafish embryo hatching. Nano‐ZnO was used as a positive control. Shedding of transition metal ions (Zn2+, Cu2+ and Ni2+) from the particle or tube surfaces were postulated to be the main reason for hatching inhibition in the embryos. Interestingly, the effective concentration of CuO (0.5 ppm) resulting in hatching inhibition is 10 times lower than ZnO (5 ppm).


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2. Based on our studies on ZnO, we hypothesize that Cu2+ and Ni2+ may also inhibit the zebrafish hatching enzyme (ZHE1) through an effect on sensitive histidines in the active enzyme center.

3. A robotic system was designed to increase the efficiency of embryo plating. Such a system is constituted with three major components, a vision recognition system, a robotic arm and a liquid dispensing system.

4. For automated data acquisition, we have successfully developed protocols of using ImageXpress and Acumen (high throughput imaging devices) to conduct automated imaging on zebrafish embryos in multi‐well plates.

IRG 5‐8: 1. Completed optimization and statistical analysis of the HTS growth inhibition assay with bacterial

knock‐out mutants using amino‐charged polystyrene (PS‐NH2) 62 nm NMs as positive controls. A full‐genome screen was performed and hits confirmed using secondary assays. 80.3% of the mutants selected as sensitive from the HTS assay with 100 mg of PS‐NH2/l and 60.7% selected as resistant from the HTS assay with 220 mg of PS‐NH2/l, were confirmed.

2. Completed bioinformatic analysis of genes controlling response to PS‐NH2. No clear metabolic pathways could be highlighted among 17 genes the absence of which resulted in resistant phenotype. Most of the 217 breakout genes resulted in highly sensitive phenotype were classified as being responsible for cell envelope biogenesis and functionality of outer membrane. Twenty of the confirmed sensitive “hit genes” had orthologs in yeast (S. cerevisiae), 16 in zebrafish (D. rerio), 27 in human and other mammals. The highest percentage of these genes were connected with energy and electron transport chain, some with DNA repair and stress. However, none of the bacterial genes having orthologs in other organisms were among those which resulted in bacterial phenotypes with highest sensitivities. Thus, the most remarkable effect of PS‐NH2 NMs towards E. coli cells was apparently specific to its cell wall structure.

3. Determine mechanism of toxicity of PS‐NH2: Comparison of confirmed sensitive “hit genes” with previous studies on antimicrobial compounds revealed these genes to be similar to those that increased the sensitivity of E. coli towards a cationic peptide. Therefore, the effect of PS‐ NH2 NMs for E. coli was hypothesized to be mediated by its positive charge. Surprisingly, no effect on growth inhibition was measured with SiO2 and its amino‐functionalized analogue and TiO2 and its amino‐functionalized analogue (the functionalized analogues were prepared by Courtney Thomas and Zongxi Li, IRG1, Dr. Jeffrey Zink group). Together with our previous results showing no toxicity of PS‐NH2 with larger original size – 162 or 600 nm, it could be suggested that the density or localization of amino group on the surface of the NMs may play an important role in their toxicity.

4. Toxicity tests with WT E.coli using additional NMs. Potential toxicity of ZnO, TiO2, CeO2, SiO2 and Ag eNMs were tested. No toxicity was seen with CeO2 (25 nm), SiO2 (28 nm) or TiO2 (25 nm) under dark conditions. Toxicity was observed with TiO2 exposed to UVB, but not UVA. Increased doping of TiO2 with Fe decreased its toxicity under UV‐B light. ZnO and Ag from the CEIN libraries assembled by IRG6 proved toxic to E. coli non‐mutated strain. Ag with primary size of 20 nm (Biopure, stabilized aqueous dispersion) was more toxic than Ag with primary size of 40 nm (Biopure, stabilized aqueous dispersion). The fact that 20 nm Ag NMs were more toxic compared to 40 nm Ag of the same structure may show their different mechanisms of action towards bacterial cells and additional analysis of 10 nm Ag toxicity towards E. coli cells may be useful.

5. Pilot‐scale HTS growth inhibition analysis with E.coli knock‐out mutants using ZnO and Ag NMs. Pilot‐scale experiments were carried out using a subset of the E.coli gene knock‐out mutant library Thirty‐six mutants were identified that displayed increased sensitivity to Ag and 10 mutants identified for ZnO NMs. The general pattern of “hit genes” was different from those observed in case of PS‐NH2 NMs. These studies will be extended for the whole E.coli knock‐out library.


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Impacts on the Overall Goals of the Center: High throughput screening projects supported by IRG5 provide a wealth of insight into nanomaterial properties associated with cellular toxicity, and provide mechanistic insight into such toxicity. Assays involving animal cells (IRG 5‐2 and 2‐1) probe a variety of biological responses but utilize a similar panel of cell types and nanomaterials, thus providing greater analytical power by enabling comparison of results. IRG 5‐2 determines sub‐lethal changes in gene transcription as a measure of stress and toxicity, while IRG 2‐1 leverages a series of hierarchical oxidative response assays developed by Nel. Findings from earlier cell‐based studies have been utilized to develop and test hypotheses in intact animals (zebrafish), thus validating cell‐based assays and accelerating discovery of mechanisms of nanotoxicity. These efforts have now led to zebrafish‐based HTS efforts (5‐6) as well as the development of hazard ranking heat maps, self‐organizing maps, and generation of QSARS by IRG 6. Assays that measure responses of single‐cell eukaryotic (yeast) and prokaryotic (bacteria) organisms both take advantage of cutting edge genomic approaches enabled by HTS capabilities of the MSSR. As initial proof‐of‐principle, a toxic positively charged eNM (PS‐NH2) was screened against libraries of both E. coli and S. cerevisiae in which every non‐essential gene is individually knocked out. These assays provided information on toxicity associated with NM properties (composition, size, dose, etc.) and also provide direct mechanistic insight into biological pathways involved in the cellular response and induction of toxicity (i.e. cell wall integrity important for prokaryotes that lack endocytosis, but a role for endocytosis genes in eukaryotic yeast). Finally, HTS and HCS assays are necessary to generate the volume of data needed by IRG6 to build models of how and why NMs display cytotoxicity. Establishment of these assays enables increased throughput with new NMs (see planned activities) that are required to generate data sufficient to begin modeling. A major positive impact for IRG5 in the current project period has been the efforts of Dr. Godwin to coordinate methods for HTS and NM dispersion as well as statistical validation of HTS. These efforts will have significant effect on the CEIN as additional experimental systems are adapted to HTS. Further, by implementing rigorous characterization of dispersion protocols, data are more readily analyzed/modeled by IRG6. Major planned activities for the Next Year: As part of the annual review of the Center research priorities, IRG 5 Leader Kenneth Bradley requested that IRG 5 be reorganized to reflect his intellectual interest for the upcoming year. While IRG5 has made an important contribution under the leadership of Dr. Bradley, he has requested that his major interest pertains to the science of high throughput screening a rather than the technicality and complexity of nanotechnology in which he has no official training. He is very interested, however, is continuing to play an active role as the faculty director of the Molecular Shared Screening Resource (MSSR) while the specifics of nanomaterials screening is handled under an IRG leader that is schooled in nanotechnology. Thus, under the proposed reorganization, Dr. Bradley will serve as the Technical Director of the Molecular Shared Screening Resource, which will assume a core function role within the Center. Dr. Robert Damoiseaux will continue to assist Dr. Bradley by providing technical consultation and assistance in the planning of high throughput experiments, including the translation of assays to HT capabilities. Scientific leadership of IRG 5 will be transferred to Dr. Andre Nel, who is proficient in mammalian high throughput screening and will be assisted by doctors Hillary Godwin and Dr. Ken Bradley in continuation and expanding bacterial high throughput screening.


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In the coming year, IRG 5 activities will focus on:

Design, acquire and integrate a robotic system that can automatically select, pick and place zebrafish embryos into multi‐well plates for screening.

Conduct studies on the mechanism of toxicity of selected NPs identified during zebrafish screening and correlating the physicochemical properties of NPs to their toxicity.

Expand the animal models for nanoparticle toxicity screening by introducing marine organisms, such as sea urchin, oyster, abalone and mussel embryos.

Expand HTS to include sub‐lethal mechanisms of toxicity in prokaryotic organisms

Apply genome‐wide knockout library screening to additional SRMs/eNMs found to be toxic to WT E. coli.

Develop and implement HTS assays using marine microorganisms and/or cells derived from marine animals (i.e. hemocytes)


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IRG 6: Modeling of the Environmental Multimedia NM Distribution and Toxicity Faculty Investigators: Kenneth Bradley, UCLA – Assistant Professor, Microbiology Yoram Cohen, UCLA – Professor, Chemical Engineering – AREA LEAD Francesc Giralt, Universitat Rovira I Virgili – Professor, Chemical Engineering Hilary Godwin, UCLA – Professor, Environmental Health Sciences Jordi Grifoll, Universitat Rovira I Virgili – Associate Professor, Chemical Engineering Barbara Herr Harthorn, UC Santa Barbara – Associate Professor, Women’s Studies/Anthropology Patricia Holden, UC Santa Barnara – Professor, Environmental Microbiology Hunter Lenihan, UC Santa Barbara – Associate Professor, Marine Biology André Nel, UCLA – Professor, Medicine; Chief, Division of NanoMedicine Robert Rallo, UCLA/ Universitat Rovira I Virgili – Associate Professor, Chemical Engineering Donatello Telesca, UCLA ‐ Assistant Professor, Biostatistics Number of Graduate Students: 1 Number of Undergraduate Students: 1 Number of Postdoctoral Researchers: 1 Goals of IRG6: IRG 6 focuses on the development of modeling and analysis tools to assess the potential environmental transport and fate of engineered nanomaterials (eNMs), develop data‐driven models of eNM toxicity based on high throughput toxicity screening assays, and develop a decision based model to enable assessment and ranking of the potential impact of new eNMs. IRG 6 research aims to: (a) identify and quantify factors affecting the transport of nanoparticles across the relevant environmental phase boundaries, (b) assess the distribution of eNMs among environmental compartments and range of potential exposure concentrations, (c) develop and/or adapt data‐driven models and data of physicochemical and toxicological properties of NMs, and (d) incorporate information from (a)‐(c) into a decision tool that will enable allow tool for hazard ranking of eNMs. Organization and integration of IRG6 Projects: Core Function:

IRG 6‐1/IRG 6‐2 ‐ Data management and Collaborative Infrastructure Current IRG 6 Research Projects:

IRG 6‐3/IRG 6‐4 ‐ Machine Learning Analysis and Modeling of High Throughput Screening Data for Nanoparticles

IRG 6‐5/IRG 6‐6 ‐ Modeling of the Environmental Multimedia Distribution of Nanoparticles

IRG 6‐7 ‐ Environmental Decision Analysis for Nanoparticles IRG 6 is pursuing its mission with the following major objectives: IRG6 consists of seven sub‐projects with the first two (IRG6‐1/IRG6‐2) being core CEIN support function. A core structure of the CEIN research collaborative system and data repository/management systems (hardware/software) repository was developed in IRG6‐1/IRG6‐2, with routine maintenance functions being the responsibility of IRG6. In order to facilitate data archiving, searching and sharing, a web‐based data management software system has been developed and deployed with continuing refinements in response to the needs of CEIN researchers. In order to address objectives (c) and (d), IRG6 has been developing the algorithmic theoretical principles and software building blocks (IRG6‐3 and IRG6‐4, respectively) in


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addition to web‐based data analysis tools necessary for knowledge extraction (i.e., data‐mining, clustering and pattern recognition) from HTS data, as well as selection and ranking of NP properties for nano‐SAR development. The above data analysis tools interface with the CEIN data management system (IRG6‐1/IRG6‐2) for direct web‐based HTS data analysis and nano‐SAR development. IRG6‐5 and IRG‐6 focus is on building modeling frameworks and associated software, respectively, for assessing the potential distribution and exposure concentrations of nanoparticles in the environment. IRG6‐7 aims to develop a web‐based decision tool to enable hazard ranking of the potential impact of eNMs, making use of information developed in IRG6‐3/IRG6‐4 and IRG6‐5/IRG6‐6, along with CEIN and other data sources. Major accomplishments of IRG 6 (Since March 2010): IRG6‐1 and IRG6‐2 (CORE CEIN SUPPORT): Data management and Collaborative Infrastructure IRG6‐1 provides CEIN core support for hardware/system software for data management, storage and computational needs of the CEIN. IRG6‐2 provides core support for data management that consists of development and maintenance/support of a web‐based data repository/management software system for the CEIN. This year the RocksCluster version 5.2 operating system and software of the CEIN cluster and system software (NAGIOS and GANGLIA) were installed and configured for automated and web‐based monitoring and supervision of all CEIN critical computation infrastructure. The CEIN infrastructure computational/data management including its server, four high‐end workstations and backup equipment is maintained by IRG6. The system is configured as an expandable cluster (presently consisting of 48 CPUs) which uses Oracle’s GridEngine for job scheduling. Two web‐based applications for high volume data management have been installed on the CEIN computer cluster for the storage and management of HTS data files. The development and maintenance of a multidisciplinary collaboration infrastructure and data management system are of vital importance for the CEIN. The data management and collaboration system are based on the Microsoft SharePoint Server. The CEIN Data Management server (CDM), based on the Microsoft SharePoint server platform, now hosts all CEIN research group sites, as well as a Data Repository site for the sharing of files/data among internal and external groups. The site now allows individual users and groups (CEIN and external) to build their own sub‐sites with document and appropriate security measures. A protocol for document (data and metadata) submission/uploading has been implemented with respect to the CEIN Data Repository and an advanced search capability is now in place. Information regarding the CDM system features and usage has been made available via video instructions, help files, online help and a periodic Newsletter. IRG6‐3/IRG6‐4: Machine Learning Analysis and Modeling of High Throughput Screening Data for Nanoparticles Efforts in this project focus on knowledge extraction from high throughput screening data of nanoparticles toxicity, development of predictive nano‐quantitative‐structure‐activity relations (nano‐SARs), feature selection for nano‐SAR development and identification of pathway linkages. Based on newly developed feature selection approach, a classification based nano‐SAR was developed for cytotoxicity of metal and metal oxide nanoparticles. This work revealed that atomization energy of the metal oxide, period of the nanoparticle metal, nanoparticle volume fraction (in solution), and the primary nanoparticle size were fundamental descriptors that enabled correlation of cytotoxicity at high level of accuracy without false negatives. The classification based nano‐SAR enables one to identify decision boundaries which are crucial for use in hazard ranking of nanoparticles. Another important development is the introduction of the self‐organizing map (SOM) analysis of HTS toxicity data. The approach was applied to toxicity data generated by IRG‐2 and IRG‐5 for cluster identification revealing


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two cluster groups corresponding to (i) sub‐lethal pro‐inflammatory responses to Al2O3, Au, Ag, SiO2 nanoparticles possibly related to ROS generation, and (ii) lethal genotoxic responses due to exposure to ZnO and Pt nanoparticles at a concentration range of 25 µg/mL‐100 µg/mL at 12 h exposure. The SOM approach has also been used to identify clusters in metal and metal oxide nanoparticles HTS cytotoxicity data. Complex network theory methods have been applied to identify relationships between cell responses (pathway activation and cytotoxicity parameters) as well as physicochemical properties of nanoparticles. Preliminary results using this approach to study the relationships between signaling pathways and observed toxicity identified the presence of three well‐differentiated communities (clusters). The use of association rules has also been adapted to identify significant relationships among the signaling pathways and the cytotoxicity. This analysis based on both RAW and BEAS‐2B mammalian cell lines, for example, indicates the presence of a hierarchical activity‐activity pattern where sub‐lethal effects such as ROS generation and the intracellular Ca2+ flux are strongly related to lethal effects (e.g. cell membrane damage and cell death). A crucial element of HTS data analysis is hit identification with strict control of false negatives or positives and to this end a series of methods have been implemented and utilized for both nano‐SAR development and cluster analysis of HTS nanoparticle toxicity. Concerning the statistical analysis for HTS data, a number of normalization and hit detection methods [e.g., Z‐score, robust Z‐score, and B‐score techniques, Signal to Background ratio, Signal to Noise ratio, Z‐factor, Z'‐factor, and Strictly Standardized Mean Difference (SSMD)] were assessed and have been implemented as (java) standalone package which will serve as a fundamental infrastructure (component) of IRG6’s web based platform for HTS data management and analysis. Tools for HTS data preprocessing (implemented in Java) and visualization (implemented in Visual Studio) have been integrated as a webpart within the architecture of the CEIN Data Management System. The webpart provides a set of techniques for array data normalization as well as for the automated hit identification. The preprocessing algorithms included in the tool are Z‐score, robust Z‐score, B‐score, Signal to Background ratio, Signal to Noise ratio, Z‐factor, Z'‐factor, and the Strictly Standardized Mean Difference (SSMD). The system is designed to automatically extract and process the HTS plate data uploaded to the CDM data repository. In addition, a classification nano‐SAR for the toxicity of metal oxide nanoparticles has been implemented via Rapidminer. The learning process provides a graphical interface for virtual (i.e. in silico) screening of nanoparticle toxicity. The application includes a validation system to assess whether or not the target NP belongs to the application domain in which the model has been developed. Finally, software has been implemented and validated for Matlab‐based implementation of the Self‐Organizing Map (SOM) algorithm. Current efforts are underway to the SOM software with the HTS data sets stored in the CDM data repository. IRG6‐5/IRG6‐6: Modeling of the Environmental Multimedia Distribution of Nanoparticles In order to develop software tools for the assessment of the environmental multimedia distribution of eNMs, a framework for model building was established. Accordingly, along with IRG4, intermedia transport pathways and associated predictive mechanistic and empirical model equations for nanoparticle intermedia transport have been compiled and are undergoing evaluation for incorporation into a multimedia modeling scheme to enable one to explore “What if?” scenarios. A first generation modeling framework, developed to predict the transport of nanoparticles, was successfully evaluated for the partitioning of TiO2 nanoparticles between air, water and soil, using basic intermedia transport processes such as dry/wet deposition and sedimentation. In addition, a library of environmental scenarios has been developed to encompass the range of potential scenarios to allow rapid analysis of the dynamic distribution (and persistence) of nanoparticles in the environment. Quantification of the


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fate and transport of nanoparticles requires information regarding their size distribution. Accordingly, a predictive computational ‘constant number’ Monte Carlo model (considering DLVO theory and accounting for sedimentation) was developed to model nanoparticle aggregation, and to determine the stable particle size distribution under various environmental conditions (e.g. pH, ionic strength, etc.). The model performance was successfully tested against experimental DLS data developed by the UCLA Water Technology Research Center and results from analytical solutions for simple aggregating systems. Improvements to the model code were implemented in order to reduce computational time and increase model accuracy when simulating a large number of particles (up to 20,000). This was necessary in order to enable the development of parameterized models (i.e., empirical models based on simulation data), based on simulations for a wide range of conditions, for use in transport and fate analysis and hazard ranking. Finally, the implementation of the fate and transport modeling efforts (IRG6‐5) into user‐friendly software (web‐based) is an ongoing effort in IRG6‐4. To date, an object oriented structure has been adopted for the development of a Multimedia Environmental Distribution of Nanoparticles (Mend‐Nano) model that will enable web‐based model execution. Software development will accelerate once the final versions of the models of IRG6‐5 are completed. IRG6‐7: Environmental Decision Analysis for Nanoparticles Rapid growth of the nanotechnology industry necessitates the introduction of decision tools to assess the potential impact of nanomaterials and options for their safe design. To date, an initial process for the decision analysis path with respect to the potential impact of nanoparticles was formulated with the goal of establishing a web‐based tool for nanoparticle impact analysis. A first version decision tree was formulated which will make use of quantitative and qualitative information provided by available data and models (including nano‐SARs developed by IRG6‐3/IRG6‐4) as well as fate and transport model developed in IRG6‐5/IRG‐6. A web‐based tool is under development based on the decision tree to enable building specific decision path scenarios. Impacts on the overall goals of the Center IRG6‐1/IRG6‐2 provide core support for the CEIN collaboration, data management and computational infrastructure. These are important CEIN functions that provide collaborative computer cluster/software environment and personal support for sharing data, documents and in an efficient workflow mode. IRG6‐1/IRG6‐2 provide the basic infrastructure required for the CEIN Data Management System. The work in IRG6‐3 on algorithms for data analysis (e.g., feature extraction, clustering, and hit identification), along with data mining tools being developed provide the algorithms/software building blocks for the development of HTS data‐driven models. Corresponding software applets developed in IRG6‐4 are being applied to CEIN data (IRG1, IRG2‐IRG‐5) to improve data analysis capabilities. Web‐based tools for HTS data analysis are now available to accelerate knowledge extraction. The NP aggregation modeling (IRG6‐5) effort has provided information on the expected size distribution of NP suspensions and will be utilized with IRG4 data to develop parameterized models that will serve other researchers to quantitatively assess the expected nanoparticle aggregate sizes under a range of conditions of environmental interest; this information is also crucial for the design of NP experimental protocols (for both physicochemical properties and toxicity) and for assessing the relative importance of transport pathways pertinent for modeling the transport and fate of nanoparticles. Efforts on the development of a decision analysis tool for hazard ranking (HR) of the potential environmental impact of nanomaterials will benefit life cycle analysis efforts by IRG4/IRG7 and will be coordinated with these activities. Finally, it is noted that the relevant intermedia NP transport pathways (IRG4) and associated predictive (and correlative) relations (IRG6‐5), along with aggregation modeling information (and CEIN data; IRG4, IRG1), are the necessary foundation for the construction of a multimedia model (IRG6‐5) and an HR based decision tool (IRG6‐7).


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Major planned activities for the next year During the next year IRG6 will focus its activities on: (i) Integration of HTS data analysis tools with the CEIN Data Management System; (ii) Collaboration with IRGs 2 and 5 to generate targeted HTS data for expansion of nano‐QSAR models utilizing the tools already developed in IRG6‐3/IRG6‐4; (iii) Continued application of network analysis in collaboration with IRG5; (iv) development of a web‐based transport and fate model for nanoparticles and incorporation of pertinent correlations/intermedia transport and physicochemical predictors into the web‐based modeling interface; (vi) Development of a hazard/risk scoring methodology for nanomaterials as web‐based platform for construction/evaluation of various decision pathway scenarios.


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IRG 7: Risk Perception of Potential Environmental Impacts of Nanotechnology Faculty Investigators: William Freudenberg, UC Santa Barbara – Professor, Environmental Studies and Sociology Barbara Herr Harthorn, UC Santa Barbara – Associate Professor, Women’s Studies/Anthropology – AREA LEAD Patricia Holden, UC Santa Barbara – Professor, Environmental Microbiology Milind Kandlikar, University of British Columbia – Assistant Professor, Institute for Global Issues Nick Pidgeon, Cardiff University – Professor, Applied Psychology Theresa Satterfield, University of British Columbia – Associate Professor, Institute of Resources Number of Graduate Students: 4 Number of Postdoctoral Researchers: 2 Goals of IRG 7: Overall: IRG 7 aims to produce new knowledge about key factors likely to drive critical stakeholder groups’ perceptions of risks to the environment posed by specific ENMs and their enabled products. We hope to use this knowledge to inform and help calibrate UC CEIN research, and public and policy outreach strategies, including recommendations to US regulatory agents. Groups/sectors to be studied include US publics, the ENM industry, and a range of experts (nanoscale scientists & engineers (NSE), nanotoxicologists, regulators, and nano environmental remediation experts). Factors currently being analyzed as drivers of public perception include environmental values, different framings of ENM environmental risks and benefits, and perceived thresholds of under‐regulation or over‐regulation by government or industry of emerging environmental risks. IRG 7 also analyzes how industry’s perceptions of risk and regulation impact their environmental stewardship & workplace safety practices and their receptivity to the regulation of engineered nanomaterials. Downstream effects of ENM production are also examined by studying how nano environmental remediation is or should be handled from the perspective of industry and regulators, and which assessment and regulatory challenges with regard to ENMs are viewed as central. The resulting portfolio of environmental risk perception research will provide UC CEIN with a carefully nuanced analysis of emergent public, industry, and expert concerns; differences among risk views of experts (those developing ENMs and their applications; those studying their risks and toxicological signature; and those responsible for their regulation); and differences among multi‐stakeholder views that could lead to conflict or controversy if not addressed upstream. IRG 7 research is developed in close consultation and collaboration with UC CEIN toxicologists and ecologists and aims to share results with the UC CEIN to contribute to responsible decision making and to increase sensitivity of UC CEIN to societal environmental concerns and values. Organization and Integration of IRG 7 Projects: IRG 7 effort is organized into 4 projects; until Dec 2010 and the untimely death of PI Freudenburg, each was led by a different lead PI (Freudenburg & Harthorn at UCSB; Satterfield & Kandlikar at UBC). Harthorn has taken over leadership of Freudenburg’s project, and a suitable new direction has been formulated, consistent with the overall IRG 7 goals and fulfilling commitments made to IRG 7 GSR Collins. Integration of IRG 7 projects is accomplished through a variety of formal and informal mechanisms, including: frequent communication among project leaders by electronic and teleconferencing means; frequent face to face meetings among UCSB personnel through workspace proximity in the CNS and UBC personnel through workspace proximity in IRES, frequent project meetings, and through regular UCSB CEIN meetings; presentations and discussions of project research methods and findings in regular meetings of UCSB CEIN, retreats, the annual ICEIN conference, and NSF


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site visits; joint participation in UC CEIN and the funder reporting mechanisms; and sharepoint data and reports sharing. IRG 7 project leaders all have additional duties in the CNS‐UCSB, and CNS‐funded IRG 3 (CNS risk perception group) meetings contribute to CEIN project integration as well. In distinct contrast to CNS risk perception research by the group, CEIN projects all have an explicit focus on environmental risk and ENMs and are clearly distinguished. CNS provides housing and infrastructure support to all UCSB‐based IRG 7 researchers. Current IRG 7 Research Projects:

IRG 7‐1: Environmental Sociology of ENMs (William Freudenburg (through Dec 2010)/Barbara Herr Harthorn (beginning Jan 2011)

IRG 7‐2: Environmental Risk Assessment and Nanomaterial Regulation (Milind Kandlikar)

IRG 7‐3: Environmental Risk Management and Regulation in the International Nanomaterials Industry (Barbara Herr Harthorn and Patricia Holden)

IRG 7‐4: Environmental Risk Perception (Theresa Satterfield and Barbara Herr Harthorn) Major Accomplishments of IRG 7 (since March 2010): IRG 7 research is advancing well in expected directions, with timelines consistent with the methods and practices of successful social science research projects.

IRG 7‐1 has completed three linked studies in the environmental sociology of ENMs, and has initiated background work for a 4th, despite significant unavoidable obstacles. The first study, currently a paper undergoing revision for resubmission to a leading journal, compares amplification potentials of early nanotech environmental risk communication in the print media with that of the early days of nuclear technology and determines that nanotech environmental risk communication has been more balanced, less benefit centric, a path CEIN is strongly recommended to follow to avoid excessive amplification in the event of possible future ENM environmental risk findings. The 2nd, also undergoing revision, looks at adverse effects on public perceptions of environmental risk of public‐private partnerships (e.g., academic and industry) at different stages of R&D (and the government) should use caution in forming industry partnerships that could lead to a perception of compromised risk management. The third (in press) is an important theoretical contribution on potential risks to the environmentally safe development of nanotechnologies of institutional and organizational failures (i.e., “recreancy”) that have ignited past environmental controversies and eroded public trust. The study serves as an important reminder that CEIN needs to be a responsible institution itself in meeting the public’s trust through transparent and timely disclosure of risks, in accessible language. A new fourth study explores potential environmental equity issues in current nanoremediation efforts by the EPA in the US that could directly impact public perceptions of UC CEIN environmental risk research. IRG 7‐1 former leader Freudenburg was honored with a lifetime achievement award from the Rural Sociology Society at their annual meetings in Atlanta, GA Aug 2010.

IRG 7‐2 has advanced its ENM regulatory life cycle analysis by completion and revision of an invited lengthy report for the Chemical Heritage Foundation (Beaudrie 2010). In summer 2010, the team completed a web survey of 424 nano S&E, nanotoxicology, and regulator experts on their views of ENM risks and regulation. This research will provide a vital comparative framework for UC CEIN public and industry risk perception studies (IRG 7‐3, IRG 7‐4). Analyses of particular likely interest to other UC CEIN researchers: general trend and agreement across experts in relative ranking of


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potential risks, with environmental releases from production facilities and occupational exposures deemed most risky compared to other release scenarios and specific nano‐applications; small but consistent differences in risk judgment across expert categories and expert gender/race; regulatory agency experts judge risks across application categories to be significantly higher than do NSE and nano EHS researchers and agree that US regulatory agencies are inadequately prepared to control risks. Study supports need for development decision‐analytic tools (risk‐ranking, multi‐criteria decision analysis, and control banding) adapted to decision making re: environmental risks of ENMs (see Beaudrie & Kandlikar, JNR 2011). To that end, IRG 7‐2 has begun plans for a nanomaterial risk ranking project (re: risk assessment under conditions of uncertainty across product lifecycles) and will explore intersections with IRG 6. Care will be taken to avoid any overlap of personnel or mission with CEIN SNUR workshop in development. CNS has agreed to co‐fund this workshop.

IRG 7‐3 has collected a novel set of data on environmental stewardship and their closely linked workplace safety practices from an international sample of ENM businesses, in addition to eliciting a set of judgments from them about the risks of particular ENMs and attitudes about their regulation. These data permit examination of the links between industry characteristics (e.g., company size, ENMs handled), their leaders’ attitudes re: risk and regulation, and their environmental and safety practices. Government, policymaker, media, and industry interest in this study’s results is high, and their dissemination represents an opportunity for UC CEIN demonstration of commitment to responsible environmental stewardship of ENMs. To this end, the group has made 12 presentations of this CEIN project this past year to industry, state and federal government, and academic audiences, including keynote addresses by Harthorn at the NNCO Capstone Conf (Mar 2010) & NIOSH (July 2010) and by Holden at Calif. DTSC (Oct 2010). Data analysis by graduate student Engeman is nearly complete, and the group is in advanced preparation of a series of journal articles for publication, anticipating completion by June 2011. A few key findings with implications for CEIN: o Overall, current guidance documents appear to be inadequate to produce the necessary

knowledge or motivations for environmental protection by ENM firms, so the time for regulation appears to be now there is strong support from this study for UC CEIN involvement with industry and regulators. To that end, Harthorn is working w/ Director Nel and others in UC CEIN on an academia‐industry‐government workshop to be held in fall 2011.

o In spite of their high level of reporting to ‘not know’ the risks of particular ENMs, and lack of use of nano EH&S programs by a majority (54%), a majority of industry participants believe that industry can be trusted to regulate itself and that voluntary reporting programs are effective in protecting the environment and workers. These findings lend support for the need for regulatory intervention.

o A majority of firms (61%) affirm the need for more information on methods for ENM EH&S, and lack of information ranks higher than budgetary constraints as an impediment to implementing nano EH&S program there is a documented gap in knowledge that UC CEIN outreach could help meet.

o The fewer the number of employees in a firm handling ENMs, the less likely the firm is to use a key nano‐specific recommended practice (i.e., monitoring the workplace for nanoparticles). Small start ups are more likely to have fewer employees handling nanomaterials hence they should be a target of outreach activities.

IRG 7‐4 is conducting research on environmental risk perception in a dually novel area (specific ENMs as nested in distinct perceptions of different environmental media). In order to accomplish this, IRG 7‐4 has completed a two‐phase design of studying public perceptions of air, water, and soil


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alone (phase 1) and in interaction with ENMs (phase 2). We have done so using mental models interview designs (which seek lay theories of cause and effect, and lay intuitions about harm and safety). Findings from the interviews were then incorporated into a pilot survey instrument. Input from UC CEIN IRGs 1, 2, 3, and 4 was used to determine which ENMs to focus on, to ensure scientific validity of the distinctions drawn among them, and to ensure instrument conformity to ecologists’ views of environmental media. The pilot survey results are currently in the data analysis phase, with a number of key findings of relevance to CEIN. A paper on the environmental values from the mental models research is in draft form for planned submission in early Spring. Several key findings from the pilot survey on environmental risk perceptions of ENMs by a large pilot sample (n<800) of US public include: o Reporting that ENMs are present in air, soil, and/or water leads to respondents scoring the

ENMs as more difficult to detect and/or measure in the environment (i.e., to touch, feel, see, describe, measure, sample and test). Those who see ENMs as highly intangible are more likely to have higher risk ratings for some materials. Implications: The very idea of invisible and intangible materials appears to inspire some risk aversion, which might be problematic for UC CEIN science communication.

o Respondents with higher tested nanotech knowledge were slightly more accepting of specific ENMs than those with little or no knowledge of nanotechnologies, though the differences are modest. Implications: Despite the above caveats, there is reason to be optimistic about the benefits of scientific literacy from UC CEIN’s science education & communication, although previous risk controversies have indicated that the knowledge benefit only holds true for risks in the absence of controversy.

o Respondents who rated the environmental media of air, water, and soil with and without added ENMs as more resilient (i.e., recovering easily from human impacts, self‐cleaning over time, mostly pure, easy to control) also tended to see the benefits of various technologies as outweighing the risks, to accept specific nanotechnologies, and to agree with reassuring statements about environmental toxicology. Implications: Emerging UC CEIN research about the actual resilience of environmental media to recover from impacts of ENMs will be salient information in the minds of some public groups, though better demographic distinction this way needs to be developed with a larger, more representative survey.

Summary total IRG 7 accomplishments for the reporting period: IRG 7‐1—major national honor from a leading prof. society; 1 publication in press, 2 in revise and resubmit; 7 presentations IRG 7‐2—3 publications, 5 presentations, 2 educational outreach IRG 7‐3—2 publications in prep, 14 presentations, 3 public outreach, 5 to policymakers, 2 to industry IRG 7‐4—7 publications, 1 in preparation, 3 presentations, 1 public outreach event IRG 7 ‐ General—1 edited volume (ed. Harthorn & Mohr); 1 special issue of Risk Analysis in prep (eds. Pidgeon, Harthorn, Satterfield) Impacts on the Overall Goals of the Center: The mission of UC CEIN is to insure that nanotechnology is introduced in a “responsible and environmentally‐compatible manner." IRG 7 contributes to that mission through multi‐stakeholder research on ENM environmental risk perception and regulatory challenges to “responsible development” that can be incorporated to enhance the UC CEIN research decision making, education and outreach activities, and for input into regulatory policy. In addition to a growing publication profile, IRG 7 has already established an extensive record of public, scientific, social science, governmental, and


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industry presentations that signficantly extends the reach of the UC CEIN and demonstrates its attention to the concerns and views of these different stakeholders (a key aim of the NNI). In the research arena, research from IRG 7 suggests a number of approaches for UC CEIN, including the following. UC CEIN should: be attentive to avoiding institutional ‘recreancy’ regarding its own responsible development obligations; be careful not to compromise its perceived trustworthiness for unbiased environmental risk assessment and communication through direct (funded) partnerships with industry; provide balanced risk‐benefit rather than benefit‐only environmental risk communication to avoid future amplification; should anticipate and adjust for diverging perceptions of ENM risk and need for regulation across the experts involved in the UC CEIN enterprise when forming UC CEIN policy and risk assessment and communication; should anticipate US nano environmental regulators’ comparative lower confidence in the sufficiency of regulatory mechanisms for ENMs; should participate in social/decision science based risk ranking exercises that will aid in the adaptation of risk regulation mechanisms to ENMs; should anticipate industry risk attenuation effects and desires for autonomy from regulation in working toward industry participation in ENM regulatory decisions; should pursue intervention with industry EH&S knowledge gap in conjunction with regulatory advances; should consider downstream industry ENM utilization patterns in determining specific ENM characterization work; should work with regulatory agencies to devise methods for outreach to small startup firms with likely higher environmental impacts; should devise plans to address cultural values with regard to different environmental media in reporting effects of specific ENMs in those media; will need to target science educational outreach and risk communication to different audiences depending on level of concerns about intangibility of ENMs and relative resilience of the environmental media. IRG 7 has recruited a strong and diverse set of postdoctoral and predoctoral researchers and is providing strong mentorship of them as well as completing educational and training opportunities for 4 UCSB Bren master’s students who are now employed (n=2) or seeking work in environmental industry, government and NGO sectors. IRG 7 students, postdocs, and researchers contribute empirical knowledge about societal implications and contexts for the risk characterization research emerging from the other groups and contribute significantly to the diversity of the UC CEIN. Major Planned Activities for the Next Year:

IRG 7‐1 will complete revisions on 2 publications on key issues for the environmentally responsible development of nanotechnologies and (re)submit them for publication. An exploratory new study uses spatial analysis to look at the citing of current nanoremediation projects being undertaken by the EPA, examining the sociodemographic composition of local communities surrounding such sites to determine if there are patterns related to the demographic distribution of technological risks and/or benefits in an emergent application of nanotechnology.

IRG 7‐2 will complete full data analysis of expert perception web survey of nano S&E, nanotoxicologists, and regulators and will work with UC CEIN researchers to consider and share implications. Planning and implementation of a workshop on lifecycle ENM risks will be a major activity for the year.

ENM Industry Survey project (IRG 7‐3) will complete preparation and submission of planned publications by early summer 2011, concluding the main work on this project. Plans include production of a comprehensive final report on the project (including biblographic literature review, project findings, and full instrument used in the study) that can be posted to the CEIN website. Dissemination on UC CEIN’s behalf will continue at several national and international venues. Project


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leader Harthorn will work w/ Nel et al. on using IRG 7‐3 findings to assist in the proposed workshop on CNT decision making tools.

IRG 7‐4 Environmental Risk Perception Survey project plans include completion and submission of the mental models research paper (Spring 2011), completion of the stage 1 pilot study, which offers key methodological insights on mental models of the perceived risks of environmental media (Summer 2011). Planning and implementation of a stage 2 survey to a larger and more representative sample will occur, with more specific ENMs for comparative risk assessment and life cycle features (Spring‐Fall 2011). IRG 7‐4 will again confer w/ UC CEIN leaders regarding specific ENMs of greatest interest and implications of the findings for ongoing research and outreach plans.

As research data are produced and provide an empirical basis for contribution, IRG 7 seeks to enhance mechanisms to integrate these research streams and the knowledge they are generating with the other IRGs in the UC CEIN and with the Education & Outreach program. IRG leader Harthorn has consulted with PI Nel and Co‐PI Godwin regarding means to facilitate this process, and that has led to additional collaborations. Examples include: Harthorn participation in the academia‐industry‐government CNT workshop in development (Nel, lead) based on IRG 7‐3 industry survey; co‐sponsorship of J. Isaacs visit to UCSB and UCLA (Feb 2011); development of a pioneering Life Cycle Analysis for ENMs workshop at UCSB (Feb 2011). Harthorn collaboration w/ UCSB CEIN PI Keller on the environmental journalism program will lead to new opportunities for dissemination of UC CEIN research in an environmentally and socially responsive manner.


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10. Center Diversity Progress and Plans The UC CEIN is strongly committed to ensuring the cultural, gender, racial, and ethnic diversity of the UC CEIN at all levels, particularly courting active involvement of women and underrepresented minorities as UC CEIN participants. We seek to ensure the broadest diversity possible by:

Partnering with UC CEIN investigator Jorge Gardea‐Torresdey from the University of Texas, El Paso. UTEP is a Hispanic serving institution. Dr. Gardea‐Torresdey interacts with the Education and Outreach staff of the UC CEIN to develop increased training opportunities for minority undergraduates ‐ primarily through seminars and internships. UC CEIN has supported the participation of graduate students and postdocs from UTEP in our Student Postdoctoral Leadership Workshops and attendance at ICEIN 2010 and ICEIN 2011.

UCLA has partnered with the Center for Nanotechnology and Society at UC Santa Barbara to recruit social science graduate students to work on IRG 7 research. 8 social sciences graduate students (including 5 females) have participated in our industry and public perception surveys over the past year.

Provide research mentoring for undergraduate and graduate researchers through partnerships with existing REU programs at UCLA, UC Santa Barbara, and UTEP, as well as with support from the recently completed UC Nanotoxicology Research and Training Program.

Seeking partnerships with faculty in minority serving academic institutions to serve as a distribution portal of curriculum developed by the Center.

Exploring partnerships that will allow for the expansion of our online graduate course offerings to participants at other Universities on an audit basis and/or for credit basis. Our recently developed online course in Nanoecotoxicology is being pilot tested by two students from the Cento de Investigaction y de Estudios Avanzados del Instituto Politechnico Nacional (CINVESTAV) in Mexico City. Plans are to expand our course offerings to both CINVESTAV and the Instituto Nacional de Salud Publico (INSP).

Through a partnership with California TEACH, the UC CEIN is providing mentoring and teacher training experiences to undergraduate students majoring in math, science and engineering. Nearly a dozen undergraduates have undergone teacher training an orientation for participation in UC CEIN outreach events at the California Science Center, Santa Monica Library, and other Center outreach events.

Recruitment of a diverse postdoctoral researcher pool. All postdoctoral scholar positions are advertised widely and publicly to ensure the broadest applicant pool.

Incorporating job skills training into our Student and Postdoctoal Leadership workshops, which will encourage successful application of our diverse students and postdocs into careers in academia and industry.

Progress since the last period: As our Center matures, we have increased engagement from a diverse range of faculty, research staff, postdoctoral scholars, graduate students, and undergraduates in our research and education/outreach activities. We have successfully engaged a high percentage of female researchers amongst our research staff, graduate students, postdocs, and undergraduates, which is notable given the traditionally low numbers of females in the fields of science and engineering. Additionally, 71% of our graduate students and 93% of our undergraduate participants were US citizens.


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While the Center does not have influence over the recruitment of new female and/or minority faculty at our member institutions, we are proud of the strong female representation in our Center leadership, with 3 area leads serving on our Executive Committee and an additional 2 female faculty active in our IRG activities. We feel this strong representation of female faculty leadership sets a strong example to up and coming scientists. Plans for the next reporting period: Over the next year, we will continue to strengthen our education and outreach partnerships, particularly those with the California Science Center, the Santa Monica Public Library, and the Brentwood School (K‐12). We will continue to actively participate in UCLA and UCSB outreach events geared towards public and K‐12 audiences. We are also exploring opportunities for expanding our undergraduate mentoring capabilities, through stronger partnerships with existing REU programs as well as seeking supplemental funds for REU student support here at UCLA. The Center remains committed to our partnership with UC Riverside (a minority serving institution) and the University of El Paso, Texas (a minority serving institution) and will explore avenues through existing and new programs to strengthen the path to higher education opportunities for minorities and women in the field of environmental nanotechnology. To help foster academic advancement for female faculty members, we are hosting a week‐long "Nanotoxicology Bootcamp" at UCLA in August 2011 to provide capacity building for researchers at Mexican higher education institutions and to promote collaborations between women scientists in Mexico and women scientists in the United States. We have recruited a diverse External Science Advisory Committee who will provide us valuable input on the Center's outreach and diversity goals.


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11. Education and Outreach Summary of Education and Outreach Goals The Education and Outreach division of the UC CEIN fosters cross‐IRG interaction and communicates Center research to industry, policy makers, the K‐12 community and the public. In order to accomplish this, our activities are focused on:

1) Fostering an interactive research environment among UC CEIN participants; 2) Co‐organizing an Annual International Meetings with CEINT; 3) Developing courses and learning modules related to nanotechnology and the environment and to

the safe handling of nanomaterials, which include content generated from the research within the Center and make content available to other institutions;

4) Implementing a Graduate Student/Postdoctoral Fellow Professional Development program; 5) Creating a family of web‐based survey tools that will be used to assess the Center, Center

Leadership, and the effectiveness of UC‐CEIN Education/Outreach; 6) Sponsoring seminars on the environmental impacts of nanotechnology in partnership with the

CNSI at UCLA and making these available to CEIN members and member institutions in real‐time via webcasting and in the CEIN’s digital archives;

7) Partnering with museums, schools, libraries, and other research centers, both in Los Angeles and in Santa Barbara, to enhance public science outreach initiatives; and,

8) Continuing to work with legislators/policy makers to ensure future legislation is based on sound science.

Summary of Education/Outreach Activities Year Three Monthly Protocols Working Group (PWG) meetings and IRG 5/HTS meetings January‐April 2010: Partnered with the California Science Center to train CEIN Volunteer

Educators and Museum Educators on nanoscience and leading NanoDays 2010 table top activities; NanoDays 2010 on April 3 at the California Science Center in Los Angeles.

March & April 2010: Partnered with the Santa Barbara Museum of Natural History and the Center for Nanotechnology in Society for NanoDays 2010 on April 3 at the SB Museum of Natural History.

April 24, 2010: Public outreach events at Santa Monica Public Library, “Nanotechnology: Small is Big!,” and Santa Barbara Public Library, “Nanotechnology: Small things on a big planet”

April 2010: Presentation Skills Workshop for graduate students/postdocs May 2010: Pre‐ICEIN 2010 Student/Postdoc Leadership Workshop May 2010: Co‐organized and co‐sponsored, with CEINT, second annual international meeting

(ICEIN 2010) at UCLA. Spring Semester 2010: Broadcast and recorded Roger Nisbet’s short course, “Dynamic Energy

Budget Theory” from UCSB; All five lectures now publicly available on UCLA iTunesU. June 2010: Carbon nanotube (CNT) discussion and interactive activity with ArtSci high‐school

summer program. October 2010: Co‐sponsored “NanoVI: Progress in Protection” symposium with CA Department of

Toxic Substances Control (CA DTSC) at UCLA. November 2010: Participated in UCLA‐based public outreach event, “Explore your universe!” To

prepare for this event, there was a lunchtime prep session for volunteers. December 2010: Student/Postdoc Advisory Committee (SPAC) conference call, planning for next

six months. January 2011: Conducted CEIN Leadership survey. January – May 2011: Launched web‐based Nanoecotoxicology lecture series for students at


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CINVESTAV and INSP in Mexico. February 2011: Initiated annual CEIN Center‐wide survey. February 4, 2011: Journalist Scientist Communication Workshop at UCLA, for UC‐CEIN

students/postdocs March 2011: CEIN Center‐wide retreat and CEIN Student/Postdoc Leadership Workshop to be held

at Lake Arrowhead, CA. March & April 2011: Partnering with the California Science Center to train CEIN Volunteer

Educators and Museum Educators on nanoscience and leading NanoDays 2011 table top activities; NanoDays 2011 on April 2 at the California Science Center in Los Angeles.

March & April 2011: Partnering with the Santa Barbara Museum of Natural History and the Center for Nanotechnology in Society for NanoDays 2011 on April 2 at the SB Museum of Natural History.

April 16, 2011: Public outreach event planned at Santa Monica Public Library, “Nanotechnology: Small is Big!”

AY 2010‐2011: Offered a three‐lecture series to Brentwood School’s AP Biology Class: Introduction to nanoscience; Nanoscience and the environment; Nanomedicine.

Sponsoring 7 seminar speakers (for joint seminars with CNSI at UCLA) at UCLA and 3 at UCSB. Elluminate Elluminate is a web‐based learning platform that allows users to participate in online courses, meeetings, or seminars. Elluminate is user‐friendly and requires that participants have only a computer with an internet connection and a telephone. The CEIN uses the Elluminate platform to hold meetings (Protocols Working Group; IRG 5/High Throughput Screening Group; ESAC) where documents are shared, to broadcast seminars and courses from UCLA or UCSB to CEIN members worldwide, and to broadcast faculty talks to external audiences when scheduling conflicts preclude travel. As CEIN's Elluminate Systems Administrator, Katy Nameth sets up and moderates each online meeting, is responsible for training graduate students to moderate sessions, and attends monthly meetings to stay up‐to‐date on systems upgrades and best practices. Principles in Nanoecotoxicology (Online Course) The UC CEIN has developed a 13 lecture online course in the Principles of the Environmental Implications of Nanotechnology. The course provides an introduction into the multidisciplinary research of the Center, including presentations on nanomaterials in regards to: manufacturing; physicochemical properties; fate and transport in the environment; impact on cells, organisms, populations, and stability of ecosystems; and tools used to assess and reduce biological harm. The course will be available to all Center members and Center educational partners. The course is currently being pilot tested by students and faculty at CINVESTAV in Mexico City, and in Summer 2011, we will run a boot camp to foster capacity building allowing researchers in Mexico to develop the knowledge basis necessary to implement and evaluate this course in their institutions. California Teach (Partner) The California Teach program encourages undergraduate students majoring in math, science, and engineering to consider a career in teaching, and participating students attend a series of teaching seminars, complete (math or science) teaching internships, and receive a stipend from UCLA. UCLA California Teach students are encouraged by their Academic Coordinator to gain more teaching and presenting experience by participating in the CEIN Volunteer Educator program.


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CEIN Volunteer Educators (Program) The CEIN Volunteer Educator program allows California Teach undergraduates and CEIN‐affiliated graduate students, postdos, and faculty to participate in public outreach events by leading hands‐on nanotechnology‐focused activities and by participating in public panel discussion (postdocs/faculty). CNSI (California NanoSystems Institute) members are also encouraged to participate in CEIN outreach events. California Science Center (Partner) The CEIN has established an outreach partnership with the California Science Center, a non‐profit public museum in Los Angeles, CA. In April of each year, the museum and CEIN co‐host a NanoDays event at the museum. In 2010, Volunteer Educators and museum educators lead a series of activities from the NanoDays kit and CEIN postdocs and faculty answered public questions about nanotechnology. The 4 hour event interacted with over 500 members of the public. The second co‐sponsored NanoDays activity was planned for April 2, 2011. Additionally, the museum has recruited CEIN volunteers to provide education sessions at their annual Spring Break science camp, their home school session, and plans are underway to have CEIN faculty give seminars to members of the museum board of directors and key donors. Santa Monica Public Library (Partner) The Santa Monica Public Library hosts free public events which complement its mission of supporting an informed and educated community. In April 2010, the library hosted CEIN public outreach event "Nanotechnology: Small is Big!" which provided an opportunity for Center scientists and community members to interact through discussions, hands‐on activities, and Q&A sessions. The 2010 event was such a success that it will be repeated in April 2011. Protocols Working Group The Protocols Working Group is an interdisciplinary group tasked with establishing procedures and policies for dissemination and validation of protocols across the UC CEin. It has met monthly since October 2009, and each meeting is webcast and archived using Elluminate. After discussion and validation of Center protocols, they will be made available via the CEIN website. Postdoctoral Training Plan The postdoctoral training plan for the UC CEIN focuses on activities that will ensure coherent and effective mentoring for all Center's postdoctoral fellows. All postdoctoral researchers develop a written training plan for their research, and undergo an annual performance evaluation with their mentor. In addition to their direct mentorship, the Center provides additional career and skills development activities to supplement the educational experience of the postdoctoral trainees. Each year, a series of leadership and skills workshops are offered to postdocs and graduate students in the Center. Daylong workshops offered to Center members over the past year include: High Throughput Screening and Analysis of Large Data Sets (May 2010); Communicating your Science to the Public (Feb 2011); and the Academic Job Search (Mar 2011). Additional sessions are offered year‐round on presentation skills, report writing, and proposal writing. Education Coordinator Katy Nameth makes herself available to all postdoctoral researchers for one‐on‐one consultation for writing and presentation skills. The Center has developed an online course in Nanoecotoxicology based on the scientific integration of our Center's research. This course will be made available for all incoming postdoctoral researchers to aid in their understanding of the interdisciplinary integration of the Center's work, as well as provide key scientific background into the Center's mission.


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The Center's Student Postdoctoral Advisory Committee (SPAC) is made up of graduate student and postdoctoral researchers from each IRG. This committee provides valuable assistance in planning the outreach and supplemental training activities for our Center's postdoctoral researchers, giving them the opportunity to indicate which skills are most important for their career development. Major Planned Activities for Next Reporting Period Over the next six months, the Education & Outreach division will:

1) Continue monthly Protocols Working Group (PWG) meetings and IRG 5/HTS meetings; 2) Contribute a lecture and interactive activity to the ArtSci high‐school summer program; 3) Hold a SPAC (Student/Postdoc Advisory Committee) conference call during Summer 2011 to plan

for the next six months; 4) Make Safe Handling training modules available on the web; 5) Hold a Presentation Skills workshop for students/postdocs; 6) Work with UC CEIN faculty, students, postdocs to identify opportunities to develop short learning

modules and examples from topics related to the environmental implications of nanotechnology that can be included in existing courses;

7) Identify upcoming funding opportunities relevant to education/outreach activities; 8) Continue to work with the State of California DTSC on EH&S issues related to carbon nanotubes

and collaborate/provide guidance for industries working in the filed of nanotechnology in California;

9) Host “Nanoecotoxicology Bootcamp” at UCLA in August 2011 to provide capacity building for researchers in Mexico and to promote collaborations between women scientists in the United States and women scientists in Mexico; and,

10) Sponsor seminars in conjunction with CEIN/CNSI joint seminar series. Organization and Integration of Education/Outreach Projects The Education/Outreach Coordinator, Katy Nameth, assists Hilary Godwin, Education/Outreach Director in coordinating activities and following through on objectives in order to meet the goals set above. In addition to being the point person for Education/Outreach, Katy has taken the lead on establishing K‐12 and public outreach events and programs as well as designing and maintaining the Volunteer Educator program and the Graduate Student/Postdoctoral Fellow Professional Development Program, managing the Center’s Elluminate (webcasting) account, and transforming training modules into interactive learning modules. Impacts on the Overall Goals of the Center We have made considerable progress on a number of goals that are central to communication across the Center and have an exciting agenda of activities lined up for the upcoming months. There is a sense of community developing amongst our graduate student and postdoctoral researchers within the Center. The protocols working group and the high throughput working group involve researchers from all aspects of the Center and creates a regular forum for discussion of integrated research topics. Development of the Safe Handling of Nanomaterials training modules has captured the interest and support of the UCLA Office of Environmental Health & Safety as well as the University of California EH&S taskforce. Discussions are underway to develop a campus wide testing of these new educational modules for implementation across UC and within Industry. The Center has quickly become a valuable resource on Nano‐EHS for policy makers, federal and state regulatory and funding agencies, and industries within California. We plan on expanding our capacity to serve as a leading reference on Nano‐ EHS research at the national level.


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To gauge the value of the UC CEIN to the Nano EH&S Working group, we conducted of survey of hte 15 members, which includes employees from the California Department of Toxic Substances Control (CA DTSC). Ten respondents rated five elements on a 5.0 scale: organizational support from UC‐CEIN; student contributions to the Nano EH&S Working Group; matrix summarizing guidance recommendations; leadership provided by UC‐CEIN in Nano EH&S for the state of California; and, UC‐CEIN’s data management system. Nano EH&S Working Group members rated the first three elements as outstanding (organizational support = 4.56, student contributions = 4.8, matrix = 4.4) and the other two elements as good (leadership = 4.4, data management system = 4.5).

Six working group members participated in NanoVI: Progress in Protection in October 2010. These six respondents were asked to rate four items on a 5.0 scale: organizational support from UC‐CEIN (4.67); scope of coverage (4.5); potential practical application of the symposium (4.17); and, their overall reaction to NanoVI (4.5).

In‐person attendees for NanoVI numbered 105; in addition, 145 people attended the webcast of the symposium. Twenty‐three in‐person attendees filled out an evaluation of NanoVI; on a 5.0 scale, these respondents rated the scope of coverage as 4.4, the potential practical application of the symposium as 4.3, and their overall reaction as outstanding, or 4.6.

Table 3a: Education Program Participants - All, irrespective of citizenship - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Student Type Total

Gender Race Data

Ethnicity:Hispanic Disabled

Male Female AI/AN NH/PI B/AA W A

More thanone racereported,AI/AN,B/AA,NH/PI

More thanone racereported,

W/A

NotProvided

Enrolled in Full Degree Programs

Subtotal 85 38 47 0 0 1 49 29 1 1 4 7 0

Undergraduate 30 13 17 0 0 0 19 8 1 1 1 3 0

Master's 8 4 4 0 0 1 3 2 0 0 2 1 0

Doctoral 47 21 26 0 0 0 27 19 0 0 1 3 0

Enrolled in NSEC Degree Minors

Subtotal 0 0 0 0 0 0 0 0 0 0 0 0 0

Undergraduate 0 0 0 0 0 0 0 0 0 0 0 0 0

Master's 0 0 0 0 0 0 0 0 0 0 0 0 0

Doctoral 0 0 0 0 0 0 0 0 0 0 0 0 0

Enrolled in NSEC Certificate Programs

Subtotal 2 1 1 0 0 0 2 0 0 0 0 2 0

Undergraduate 0 0 0 0 0 0 0 0 0 0 0 0 0

Master's 0 0 0 0 0 0 0 0 0 0 0 0 0

Doctoral 2 1 1 0 0 0 2 0 0 0 0 2 0

Practitionerstaking courses 0 0 0 0 0 0 0 0 0 0 0 0 0

K-12 (Pre-college) Education

Subtotal 991 0 1 0 0 0 1 0 0 0 0 0 0

Teachers 1 0 1 0 0 0 1 0 0 0 0 0 0

Students 990 0 0 0 0 0 0 0 0 0 0 0 0

Total 1078 39 49 0 0 1 52 29 1 1 4 9 0

LEGEND:

AI/AN American Indian or Alaska Native

NH/PI Native Hawaiian or Other Pacific Islander

B/AA Black/African American

W White

A Asian, e.g., Asian Indian, Chinese, Filipino, Japanese, Korean, Vietnamese, Other Asian

More than onerace reported,AI/AN, B/AA,NH/PI

Personnel reporting a) two or more race categories and b) one or more of the reported categories includes American Indian or Alaska Native, Black orAfrican American, or Native Hawaiian or Other Pacific Islander

More than onerace reported,W/A

Personnel reporting a) both White and Asian and b) no other categories in addition to White and Asian

US/Perm U.S. citizens and legal permanent residents

Non-US Non-U.S. citizens/Non-legal permanent residents

.

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Table 3b: Education Program Participants - US Citizens and Permanent Residents - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Student Type Total

Gender Race Data


Male Female AI/AN NH/PI B/AA W A



W/A

NotProvided

Enrolled in Full Degree Programs

Subtotal 67 30 37 0 0 0 43 19 1 1 3 7 0

Undergraduate 28 12 16 0 0 0 18 7 1 1 1 3 0

Master's 7 3 4 0 0 0 3 2 0 0 2 1 0

Doctoral 32 15 17 0 0 0 22 10 0 0 0 3 0

Enrolled in NSEC Degree Minors

Subtotal 0 0 0 0 0 0 0 0 0 0 0 0 0

Undergraduate 0 0 0 0 0 0 0 0 0 0 0 0 0

Master's 0 0 0 0 0 0 0 0 0 0 0 0 0

Doctoral 0 0 0 0 0 0 0 0 0 0 0 0 0

Enrolled in NSEC Certificate Programs

Subtotal 0 0 0 0 0 0 0 0 0 0 0 0 0

Undergraduate 0 0 0 0 0 0 0 0 0 0 0 0 0

Master's 0 0 0 0 0 0 0 0 0 0 0 0 0

Doctoral 0 0 0 0 0 0 0 0 0 0 0 0 0

Practitionerstaking courses 0 0 0 0 0 0 0 0 0 0 0 0 0

Total 67 30 37 0 0 0 43 19 1 1 3 7 0

LEGEND:




W White








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12. Outreach and Knowledge Transfer One of the major goals of the Center is to train the next generation of nano‐scale scientists, engineers, and regulators to anticipate and mitigate potential future environmental hazards associated with nanotechnology, while at the same time seeking to impact the scientific, educational, and policy communities both nationally and internationally. We seek to educate the broader community through both UC CEIN sponsored seminars and workshops, as well as active participation in scientific meetings nationally and internationally across the range of UC CEIN disciplines. The Center has become a valuable resource, and our public profile as that of a leading Center for research on Nanotechnology Environmental Health and Safety continues to rise on both the local, state, national, and international level. Key outreach and knowledge transfer activities of the Center for the period April 1, 2010 ‐ March 31, 2011 include: UC CEIN Sponsored Activities UC CEIN Seminar Series

April 13, 2010 ‐ Fishing to define the nanoparticle properties that dictate biological responses ‐ Robert Tanguay, Oregon State University ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

April 26, 2010 ‐ Understanding the Biological and Environmental Implications of Nanomaterials ‐ Sijie Lin, Clemson University ‐ Held at UCLA.

May 4, 2010 ‐In Quest of a Systematic Framework for Unifying and Defining Nanoscience ‐ Donald Tomalia, Central Michigan University ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

May 5, 2010 ‐ The Legal Implications of Nanotechnology and Other Emerging Technologies ‐ Lynne Bergeson, Bergeson & Campbell Law Firm ‐ Webcast from UCSB.

May 11, 2010 ‐ Nanotechnology in the Public Eye ‐ Sharon Dunwoody, University of Wisconsin‐Madison ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

May 11, 2010 ‐ Developing Safe Nanotechnologies ‐ Are We Getting What We Ask For? ‐ Andrew Maynard, University of Michigan ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

October 12, 2010 ‐ Nanotechnologies and Nanomaterials in the Occupational Setting ‐ Paul Schulte, NIOSH ‐ CEIN/CNI Seminar Series ‐ Held at UCLA (archived on CNSI website).

November 16, 2010 ‐ Nanomaterials and Metals ‐ April Gu, Northeastern University ‐ Webcast from UCSB.

November 17, 2010 ‐ Dr. David Vandenberg, Lab Saftey and IIPP Program Manager, UCSB EH&S ‐ UCSB (hosted by CNS, co‐sponsored by CEIN).

November 22, 2010 ‐ Promise and Challenge of Metal Nanomaterial for Human Health ‐ Wentong Lu, Jackson State University ‐ Held at UCLA

December 1, 2010 ‐ Regulating Emerging Technologies to Protect Workers, Communities, and Environment ‐ Shiela Davis, Silicon Valley Toxics Coalition ‐ USCB (CNSI seminar co‐sponsored by CEIN).

December 2, 2010 ‐ Occupational and Environmental Health and Safety for Nanotechnology: Setting the Pace for the Next Phase ‐ Su‐Jung Candace Tsai, University of Massachusetts, Lowell ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

February 7, 2011 ‐ Life Cycle Assessment of Nanomaterials ‐ Daylong workshop co‐lead by Jackie Isaacs, Northeastern University and Sangwon Suh, UC Santa Barbara ‐ Webcast from UCSB


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February 9, 2011 ‐ Nanotechnology Environmental Health & Safety ‐ Jackie Isaacs, Northeastern University ‐ CNS Research Seminar, co‐sponsored by CEIN>

February 15, 2011 ‐ Use of Life Cycle Assessment Methodologies with Emerging Nano‐enabled Products ‐ Jackie Isaacs, Northeastern University ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

March 8, 2011 ‐ Cancer Nanotechnology: Opportunities and Challenges; View from the NCI Alliance for Nanotechnology in Cancer ‐ Piotr Grodzinski, National Cancer Institute ‐ CEIN/CNSI Seminar Series ‐ Held at UCLA (archived on CNSI website).

Workshops co‐sponsored by UC CEIN International Conference on Environmental Implications of Nanotechnology 2010 May 11‐13, 2010 ‐ University of California Los Angeles. Co‐sponsored by CEINT

The second joint international meeting of the NSF/EPA funded CEIN programs took place on May 11‐13, 2011 at the University of California Los Angeles California NanoSystems Institute in Los Angeles, CA. Researchers from around the globe in Environmental Nanotechnology presented findings to an audience of over 200 scientists, policy makers, and stakeholders. 48 talks and 50 posters were presented in the following research tracks:

Fate, Transport, and Transformation

Toxicity/Ecotoxicity

Risk Perception, Risk Assessment, and Life Cycle Analysis

Natural Nanomaterials and Nanobiogeochemistry

Nanomaterials Characterization and Toxicity Screening

Ecology and Ecosystems Response A detailed agenda and list of speakers is available on the CEIN website (http://www.cein.ucla.edu)

Nano 2010 August 22‐26, 2010 ‐ Clemson University. UCCEIN co‐sponsored event. (http://www.clemson.edu/public/nano2010/)

Nano 2010 provided a venue for presentation and discussion of current research, bringing together an interdisciplinary mix of environmental scientists, toxicologists, material scientists, and engineers. UC CEIN co‐sponsored this year's meeting, the 5th in an ongoing international series.

Nanotechnology VI: Progress in Protection October 13, 2010. Sponsored by: The UC Center for Environmental Implications of Nanotechnology (UC CEIN) and the California Department of Toxic Substances Control (DTSC) (from Ed‐10) co‐sponsored by the California DTSC, CNSI at UCLA, UC‐CEIN, COEH at UCLA, the Southern California Education and Research Center at UCLA and the Luskin Center for Innovation at UCLA.

Nanotech VI was a daylong workshop held in the California NanoSystems Institute of the University of California, Los Angeles on October 13, 2010. Nanotech VI built upon topics discussed in DTSC’s previous Nanotech symposia while emphasizing occupational safety and health concepts which are key in reducing potential risks posed by Engineered Nanomaterials (ENM’s) to workers and the environment. The day's events were presented under the banner:


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"Progress in Protection" Nanotech VI focused on techniques and approaches which can be used to anticipate, recognize, evaluate and control occupational exposure to ENM’s. This symposium served to further demonstrate that California is actively engaged not only with nanotechnology manufacturing, but also nanotechnology Environmental Health and Safety (EH&S) discussions and guidance. In partnership with the Southern California Education and Research Center at UCLA, we were able to provide Continuing Education Credits/Professional Certification Maintenance points through the following CE programs: ‐ American Board of Industrial Hygiene (ABIH) ‐ .92 IH Maintenance Points ‐ Board of Certified Safety Professionals (BCSP) ‐ .65 Continuation COC/CEU Points ‐ Registered Environmental Health Specialist Program (REHSP) ‐ 7 Contact Hours 105 registrants attended the symposium in person, with 145 additional participants viewing the workshop live online.

UC CEIN Santa Barbara Monthly Seminar Series UC CEIN members from UC Santa Barbara meet once a month to review progress across projects. EAch meeting, project presentations are made from select projects and a discussion is held. Presentations over the past year included:

April 15, 2010 ‐ Cassandra Engeman and Lynne Baumgartner. Industry survey, presentation of preliminary findings.

August 2010 ‐ Milka Montes. Gold nanoparticle biosynthesis and characterization.

August 31, 2010 ‐ Mary Collins. Evaluating risk perception of nanomaterials in the environment.

September 2010 ‐ Milka Montes. Fate and transport of nanoparticles in biological surfaces.

September 2010 ‐ Youn‐Joo An. Interactions between bacteria and nanoparticles.

September 2010 ‐ Seung Woo Jeong. Nanoparticles at the air‐water interface.

October 2010 ‐ Dongxu Zhou. Mechanism responsible for the initial aggregation of metal oxide nanoparticles.

October 2010 ‐ Raja Vukanti. Development of HTS‐Protocols for Nanomaterial Effects on Bacteria.

December 2010 ‐ Shannon Hanna. Effects of chronic exposure to ZnO NPs on the Mediterranean mussel Mytilus galloprovincialis.

December 2010 ‐ Yuan Ge. Evidence for Effects of TiO2 and ZnO Nanoparticles on Soil Bacterial Communities.

December 2010 ‐ Trish Holden. The Impact of Nanomaterials on Organisms.

January 2011 ‐ Konrad Kulacki. Effects of TiO2 on freshwater phytoplankton: general impacts and specific mechanisms.

January 2011 ‐ Samuel Bennett. Photodissociation of metal oxide nanoparticles.

February 2011 ‐ Reginald Thio. Nanoparticle‐Mineral Surface Interactions: Fate and Transport in the Environment.

February 2011 ‐ Milka Montes. Uptake and bioprocessing of CeO2 and ZnO by M. galloprovincialis.

Academic Courses incorporating CEIN‐related content


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Courses Made Available to UC CEIN Members via Webcast

DEB Modeling ‐ An 10 lecture short course in Dynamic Energy Budget modeling was made available to international participants in Spring 2010, taught by UC CEIN Co‐PI Roger Nisbet (UC Santa Barbara). In addition to 14 in person attendees, 60 international participants registered online for the course and over 30 participants participated in 6 or more lectures. The course has been archived and made publically available on iTunes U.

Nanoecotoxicology Principles. The Center previously offered an interdisciplinary capstone course at UCLA on the principles of Nanotoxicology, which was made available to Center members at other institutions via webcast. This year, we have revised the course and developed a 13‐lecture series that includes a large amount of content generated from our Center research. The lecture series is intended for a multi‐disciplinary audience and will be made available to all of our partner institutions via a secured website. In Winter 2011, we partnered with the Centro de Investigacion y de Estudios Avanzados del Instituto Politechnico Nacional (CINVESTAV) in Mexico to offer this course for credit to two PhD toxicology students. After this successful pilot, we are holding a week‐long "bootcamp" this summer for instructors who wish to learn about the research areas covered in the course. This capacity building exercise will allow researchers to become proficient in the material to the extent that they can execute and grade the course with minimal input from the Center.

Courses taught incorporating CEIN content (not webcast)

Fall 2010—Harthorn led Soc 591 BH‐‐CNS research seminar for grads, postdocs, NSE fellows, visitors, researchers: Theme for Fall 2010: Nanotechnology Environmental Health & Safety

NP synthesis and toxicity incorporated into core class and lectures that are part of NSF‐IGERT bridging the School of Engineering and the School of Health Sciences ‐‐‐‐‐Sandia/U New Mexico ‐

Trish Holden ‐ Organized / led new Bren course: ESM595J: Seminar in Ecotoxicology. Fall 2010. Speaker series (4 total) which included one nano‐speaker (April Gu, Elluminated) and one UC CEIN investigator (Gary Cherr) as another speaker.

Lectures, Seminars, and Presentations by UC CEIN members to external audiences Christian Beaudrie, University British Columbia

Risk Assessment and Nanomaterial Regulation, ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Risk and Nanomaterial Regulation: A Life Cycle Investigation of Federal Health and Environmental Regulations (Poster), ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Risk and Nanomaterial Regulation: A Life Cycle Investigation of Federal Health and Environmental Regulations (poster), Greener Nano 2010 Conference, Portland, Oregon, June 16, 2010.

Lessons Learned from a Survey of Nanotechnology Experts, RMES 500s, Qualitative Methods in Interdisciplinary Contexts, University of British Columbia, November 17th, 2010.

Technology and Sustainability? Institute for Resources, Environment and Sustainability Student‐Led Seminar Series, University of British Columbia, November 30th, 2010.

Benefits, Risks, and Regulation of Nanomaterials: Results from an Expert Survey, Society for Risk Analysis Annual Meeting, Salt Lake City, Utah, December 5‐8, 2010.


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Samuel Bennett, UC Santa Barbara

Photoactivated behavior of nano‐metal oxides in natural waters (poster), ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Raven Bier, UC Santa Barbara

Effects of nano‐TiO2 on the growth and metabolism of common freshwater algae (poster), ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

C. Jeffrey Brinker, University of New Mexico/Sandia

Engineered Biotic/Abiotic Materials and Interfaces for Understanding and Controlling Biology, Plenary lecture, XIX International Materials Research Congress 2010, August 15‐19, 2010, Cancun, Mexico.

Engineered Biotic/Abiotic Materials and Interfaces for Understanding and Controlling Biology, Karolinska Institutet, Swedish Medical Nanoscience Center, Sept 7, 2010, Stockholm, Sweden.

Directing Sol‐Gel Processing with Proteins and Living Cells. Robert B. Sosman Award Lecture, American Ceramics Society, Materials Science and Technology 2010 Conference, October 17‐21, 2010, Houston, TX.

Bradley Cardinale, UC Santa Barbara

TiO2 nanoparticles stimulate biomass production in freshwater algae. North American Benthological Society annual conference, Santa Fe, NM. June 6‐10, 2010.

H. Castillo ‐Michel, University of Texas El Paso

Chemical and elemental mapping and speciation of As and ZnO nanoparticles in plants using synchrotron radiation, European Synchrotron Radiation Laboratory, Grenoble, France. October 5, 2010.

Coordination and speciation of arsenic in the root‐soil interface of the desert plant Prospis juliflora Advanced Light Source User's meeting 2010, Berkeley, CA. October 13, 2010.

Indranil Chowdhury, UC Riverside

A Novel Microscope‐based Study on Deposition and Attachment Mechanisms of TiO2 Nanoparticle on Surfaces (Poster), ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

A Novel Microscope‐based Study on Deposition and Attachment Mechanisms of TiO2 Nanoparticle on Surfaces, 84th Colloid and Surface Science Symposium, American Chemical Society, June 20‐23, 2010

A Novel Microscope‐based Study on Deposition and Attachment Mechanisms of TiO2 Nanoparticle on Surfaces, presentation delivered at the Chemical and Environmental Engineering Graduate Symposium, University of California, Riverside, CA, September 2010

Deposition of TiO2 Nanoparticles on Surfaces in Parallel Plate Chamber: Role of Natural Organic Matter”, National Meeting of American Institute of Chemical Engineers, November 2010.

Fate, Transport and Removal of nano‐TiO2 in Aquatic Environment: Fundamental Mechanisms and Implications (poster), University of California Global Health Day, University of California, Irvine, CA, November 30, 2010

Mary Collins, UC Santa Barbara

Technological Risk Messages: Comparing Nuclear Power and Nanotechnology (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.


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Technological Risk Messages: Comparing Nuclear Power and Nanotechnology. AESS conference, Portland, Oregon, June 17 to 20, 2010.

Corporate Environmental Justice Performance: Place‐Based Facility Targeting Using A Disproportionality Framework. AESS conference, Portland, Oregon, June 17 to 20, 2010.

Temporal myopia: a case of new technology, the federal government and inherent conflicts of interest." Presentation at the American Sociological Association meetings, Atlanta, Aug 14, 2010

Recreancy and Nanotechnology: A Call for Empirical Research.” Paper presented at the Society for Risk Analysis, Salt Lake City, Dec. 6‐8, 2010.

Gwen D'Arcangelis, UCSB

‘Mental models’ of Environmental Risk Perception: Surveying Public Response to Nanomaterials (poster), ICEIN 2010. Los Angeles, California, May 11‐12, 2010

Public risk perception of environmental risks of ENMs and environmental justice.” Presentation at National Women’s Studies Association, Nov 12, 2010, Denver.

Guadalupe de la Rosa, University of El Paso Texas

Determination of Toxicity of CeO2 Nanoparticles on Soybean Plants grown in Hydroponics," National Association of Black Geologists and Geophysicists 29th Annual Technology Conference, San Antonio TX, 9‐10 September 2010.

Determination of the toxicity of CeO2 nanoparticles on soybean plants grown in hydroponics," Society of Environmental Toxicology and Chemistry 31st Annual Meeting, Portland OR, 10 Nov 2010.

Cassandra Engeman, UC Santa Barbara

Reported Practices and Perceived Risks Related to Health, Safety, and Environmental Stewardship in Nanomaterials Industries (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010

Current Practices and Perceived Risks Related to Health, Safety and Environmental Stewardship in Nanomaterials Industries. International Sociological Association, Gothenburg Sweden, July 13, 2010

Ellie Fairbairn, University of California Davis

Metal Oxide Nanomaterials in Seawater: Linking Physical Characteristics with Biological Response in Sea Urchin Development (Platform Presentation and Poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Environmental Implications of Nanotechnology. Sonoma State University, Department of Biology colloquium. October 26, 2010.

Metal Oxide Nanomaterials in Seawater: Linking Physical Characteristics with Biological Response in Sea Urchin Development(Platform Presentation). Society of Environmental Toxicology and Chemistry (SETAC), Annual meeting. Portland, OR, November 10, 2010.

Jorge Gardea‐Torresdey, University of Texas El Paso

My phytoremediation journey. 3rd Symposium Consortium Japan‐Mexico‐United States of Technological Engineering, University of Guanajuato, Gto., Mexico, March 8, 2010, keynote talk

Toxicity and biotransformation of Ni(OH)2 nanoparticles. University of Florida, Gainesville, FL, March 12 2010, invited seminar speaker.


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Toxicity and biotransformation of Ni(OH)2 nanoparticles. West Virginia, April 7, 2010. Invited seminar speaker.

Nanotecnologia ambiental: impacto de las nanoparticulas metalicas en plantas terrestres. X Congreso Nacional de Microscopia, Morelia, Michoacan, Mexico, May 23‐27, 2010, Invited talk.

Especiacion de cromo en biomasa de girasol mediante espectrosocpia de absorpcion de rayos X. VII Encuentro Participacion de la Mujer en la Ciencia, Leon Guanajuato, Mexico, May 26‐28, 2010.

Toxicity and biotransformation of nanoparticles on Terrestrial plants: The case of Ni(OH)2, ZnO and CeO2 nanoparticles. Department of Biotechnology, Indian Institute of Technology‐Roorkee, Roorkee, India, June 23, 2010, invited talk.

Nanoparticles and the Environment Seminar.," University of New Orleans, New Orleans LA,. August 10, 2010.

Hazards on the Biotransformation of Nanoparticles by Terrestrial Plants," Presentation at US EPA Region VI, Dallas TX. August 11, 2010.

Evidence of the differential biotransformation and genotoxicity of ZnO and CeO2 nanoparticles on soybean (Glycine max) plants.7TH International Phytotechnology Conference, University of Parma, Parma, Italy, 26‐29 September 2010.

Biotransformation of nanoparticles in the environment: Is it a hazard? CRETE 2010, 2nd

International Conference on Hazardous and Industrial Waste Management, Chania, Crete, Greece, October 5‐10, 2010.

How to get published, workshop 4th, Chania, Crete, Greece, October 8, 2010.

Toxicity and biotransformation of nanoparticles on Terrestrial plants: The case of Ni(OH)2, ZnO and CeO2 nanoparticles.” NanoScience Technology Center and Advanced Materials Processing & Analysis Center Seminar. Orlando, FL, October 29, 2010.

Seminar at the University of Texas‐Austin,, "Biotransformation of Nanoparticles on Terrestrial Plants," Department of Civil, Architectural and Environmental Engineering. November 18, 2010.

Evidence of the differential biotransformation and genotoxicity of ZnO and CeO2 nanoparticles on soybean (Glycine max) plants. SW/SE regional ACS meeting in New Orleans, LA, November 30‐December 4 2010.

Biotransformation and speciation of nanoparticles on Terrestrial plants using XAS: The case of Ni(OH)2 , ZnO and CeO2 Nanoparticles. 2° Encontro Brasileiro sobre Especiação Química (Second Brazilian Meeting on Chemical Speciation) ‐ EspeQ/Brasil/‐São Pedro, 2010 12‐15 December 2010.

Evidence of the differential biotransformation and genotoxicity of ZnO and CeO2 nanoparticles on soybean (Glycine max) plants. The 2010 International Chemical Congress of Pacific Basin Societies, Honolulu, Hawaii, December 15‐20, 2010.

Toxicity and Biotransformation of Metal Oxide Nanoparticles in the Environment. The Case of Terrestrial Plants” Department of Chemistry, Universidad de Castilla‐La Mancha, Ciudad Real, Spain, January 25, 2011.

Our Phytoremediation Journey: From Gold to Chromium” Universidad de Castilla‐La Mancha, Ciudad Real, Spain, Institute for Chemical and Environmental Technology, January 26, 2011.

Yuan Ge, UC Santa Barbara

Evidence for negative effects of TiO2 and ZnO nanoparticles on soil bacterial communities (postser). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

TiO2 and ZnO nanoparticles negatively affect soil bacterial communities” (poster), ASA‐CSSA‐SSSA 2010 International Annual Meetings in Long Beach, California, Oct 31 – Nov 3, 2010.


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Saji George, UCLA

Toxicity assessment of nanomaterials for safe nanotechnology applications. Biotoxicity Workshop: High School Nanoscience Program, CNSI, Los Angeles. March 13, 2010.

High throughput screening development in mammalian cells using macrophages and epithelial cells to develop paradigms for assessment of nanomaterial toxicity. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

A High Throughput Cytotoxicity Screening with In Vivo Relevance for Toxicity Profiling of Engineered Nanomaterials”, NANO 2010, Clemson, SC, August 2010.

Nanotechnology and its implications to human and environmental health (Invited lecture). Brentwood High School, Los Angeles, USA. October 20, 2010.

Hilary Godwin, UCLA

Nanoscience & The Environment” at Brentwood School, to AP Bio class of 30 high‐school students. January 2011

Shannon Hanna, UC Santa Barbara

Toxicity of ZnO nanoparticles to a soft‐sediment estuarine amphipod. 7th Annual EEMB Graduate Student Symposium, Santa Barbara, CA, February 12, 2011.

Jose Hernandez‐Viezcas, University of Texas El Paso

Biotransformation and distribution of ZnO and CeO2 nanoparticles in the desert plant Prosopis juliflora. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Taimur Hassan, UCLA

CEIN approach to data management and collaboration, Nanoinformatics 2010, Arlington, VA, November 3‐5, 2010.

Barbara Herr Harthorn, UC Santa Barbara

PCAST testimony on importance of public views to NNI mission, Palo Alto, Feb 2010?

NNCO Capstone meeting, Keynote address. Arlington, Virginia, March 30‐31, 2010.

Characterizing the Nano workforce. NIOSH worker safety conference, Keystone, Colorado, July 20‐22, 2010.

Deliberating Risks: Public Perceptions Regarding Nano Food and Agricultural Applications. Rural Sociological Society Annual Meeting. Atlanta, Georgia. August 12‐15, 2010.

Paradoxes of Development: Techno‐Enthusiasm and Skepticism in US Nanotech Deliberation. Society for the Study of Nanoscience and Emerging Technologies (S.NET), Darmstadt, Germany, Sept 29‐Oct 2, 2010

Ambivalence, uncertainty & risk: Public engagement with new [nano]technologies. Globalizing Risk UCSB Faculty Lecture Series, American Cultures & Global Contexts, University of California at Santa Barbara. Oct. 22, 2010.

Risk Perception and Environmental Health and Safety Practices in the Global Nanomaterials Industry,” guest lecture to UCSB Anthropology/Environmental Science 130A: Coupled Human and Natural Systems, November 10, 2010.

Nothing New about Nano? Making Interdisciplinary Advances in Risk Perception Research. Symposium Chair, Society for Risk Analysis, Salt Lake City, Dec 5‐8, 2010.

What’s New about nano? Nanotechnology risk perception specialist meeting Jan 2010. the Society for Risk Analysis, Salt Lake City, Dec 5‐8, 2010.


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New methods for public and other stakeholder participation. NNI and Societal Impact session, NSF Nanoscale Science & Engineering Granteees Conference. Arlington, VA, Dec 8, 2010.

Patricia Holden, UC Santa Barbara

Potential interactions and ecological manifestations between engineered nanomaterials and environmental bacteria. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Multidisciplinary Approaches and Insights into the Ecotoxicology of Engineered Nanomaterials by the UC CEIN. New England Nanomanufacturing Summit 2010, Lowell, MA, June 22, 2010.

Assessing Potential Ecological Implications of Engineered Nanomaterials: Insights from within the UC CEIN. Nano2010: Environmental Effects of Nanoparticles and Nanomaterials, Clemson University, S. C. August 23‐26,2010.

UCSB Nanotechnology Industry Survey Overview; Nanomaterial Eco‐Toxicology Impacts. October 13, 2010. Nanotechnology VI: Progress in Protection.

Bioavailability and Fates of CdSe and TiO2 Nanoparticles in Eukaryotes and Bacteria. U. S. EPA Nanotechnology Grantees Meeting, November 8‐9, 2010. Portland, OR.

Approaches and Rationale for Studying Fates and Effects of Engineered Nanoparticles in Bacterial Systems. November 10, 2010. SETAC North America 31st Annual Meeting, Portland, OR.

Bioavailability and Effects of Manufactured TiO2 and Quantum Dot Nanomaterials to Environmental Organisms. Fall Meeting of the American Geophysical Union, San Francisco. Session: Nanoparticles in Environmental Media II, December 13, 2010.

Allison Horst, UC Santa Barbara

Dispersion and reduced settling of initially agglomerated TiO2 nanoparticles due to association with Pseudomonas aeruginosa bacteria.” ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Tin Klanjscek, UC Santa Barbara

Towards a mechanistic model of toxicity of nanomaterials (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Angela Ivask, UCLA

Mechanism‐based profiling of toxicity of nanoparticles using a set of Escherichia coli strains: differentiating the impact of particles and solubilized metals (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Zhaoxia Ivy Ji, UCLA

Nanoparticle Dispersion and Stability Evaluation in Cell Culture Media. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Milind Kandlikar, University of British Columbia

Risk Prioritization for Regualting Nanoamaterials. Conference on Governing Nanobiotechnologies, University of Minnesota, Minneapolis, MN, April 15, 2010.

Konrad Kulacki, UCSB

Effects of nano‐TiO2 on the structure and function of stream ecosystems: An ongoing experiment in freshwater mesocosms. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.


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Effects of nano‐TiO2 on freshwater phytoplankton: General impacts and specific mechanisms. Society of Environmental Toxicology and Chemistry annual conference, Portland, OR, November 10, 2010.

Haven Liu, UCLA

Computational Model of Nanoparticle Aggregation Kinetics. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Modeling Aggregation and Size Distribution of Nanoparticles via Monte Carlo Simulations (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Modeling Aggregation and Size Distribution of Nanoparticles with Monte Carlo Simulation, 2010 NanoTech Conference, Anaheim, California, June 24th, 2010.

Rong Liu, UCLA

Nanoparticles toxicity: knowledge extraction from high‐throughput screening data. Nanoinformatics 2010, November 3‐5, 2010, Arlington, VA.

Computational Simulation of Nanoparticle Aggregation, 2010 Annual AIChE Meeting, November 7‐12, Salt Lake City, UT.

Lutz Madler, University of Bremen

Universität Duisburg, Germany, February 4, 2010

NanoKommision of the German Government, Berlin, Germany, February 17, 2010

Hanse Wissenschaftskolleg, Delmenhorst, Germany, February 18, 2010

Green nanomaterials: nanoparticles with increased biological acceptance, World Congress in Particle Technology, WCPT6 2010, April 26‐29, Nürnberg, Germany.

Universität Erlangen Nürnberg, Germany, May, 27, 2010

TU Braunschweig, Germany, June 17, 2010

Nanostructured materials through combustion synthesis, 9th International Congress of Chemical and Process Engineering CHISA 2010 and the 7th European Congress of Chemical Engineering ECCE Prague, Czech Republic, August, 28, 2010 (Opening Plenary of Conference)

Funktionelle Nanopartikel als Bausteine für hoch poröse Filme NanoMat Trend, Weinheim, Germany, September 5, 2010.

Flame made TiO2: Pt‐functionalization and the synergistic effect of anatase and rutile Internat. CECAM Workshop, Bremen, Germany, September 9, 2010.

Characterization of nano‐bio‐interactions, Columbia University New York, New York, USA, October 19, 2010

Rutgers University New York, New York, USA, October 20

Innovationen aus der Düse: Prozesse, Materialien, Produkte, Technologiepark Bremen, Bremen, Germany, November 18, 2010

Metalloxide aus der Sprühflamme: Herstellung, Charakterisierung, Anwendung, Fraunhofer Institute for Silicates Research (ISC), Würzburg, Germany, December 7, 2010

Nanostructured materials through combustion synthesis, Func‐Band workshop University of Bremen, Bremen, Germany, January 14, 2011

Randall Mielke, UC Santa Barbara

Quantum Dots Processed and Formed by Pseudomonas aeruginosa as Examined by Low‐Accelerating Voltage STEM‐EDX (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.


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Milka Montes, UC Santa Barbara

Biosynthesis of gold nanoparticles using alfalfa biomass. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Sumitra Nair, UCLA

Learning the effect of nanoparticles on biological cells using property activity relationships (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Catherine Nameth, UCLA

Rethinking your course for an online audience, @ONE Desktop Seminar (Learn‐at‐lunch series)

Engaging and Effectively Communicating our Results to a Broad Range of Stakeholders: UC CEIN Education and Outreach Activities (poster), ICEIN 2010, Los Angeles, CA, May 11 ‐ 12, 2010

Andre Nel, UCLA

UC Center for Environmental Implications of Nanotechnology. Invited Keynote Talk. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Nanotoxicology as a Predictive Science: from Cells to Whole Animals, Third Annual International NanoBio Conference, Zurich, Switzerland, August 26, 2010.

Nanotechnology Environmental, Health, and Safety Issues . WTEC, Nanotechnology Long‐term Impacts and Research Directions: 2000‐2020, Washington, DC, September 29, 2010.

Nanotechnology as a Predictive Science that can be Explored by High Content Screening and the use of Computer‐Assisted Hazard Ranking. Nanoinformatics 2010, Arlington, VA, November 3, 2010.

Nanotechnology as a Predictive Science: From Cells to Whole Animals. NIEHS Keystone Science Lecture Series, Research Triangle Park, NC, November 17, 2010.

Nano2: Long‐term View for Nanotechnology EHS. NSF Nanoscale Science and Engineering Conference, Arlington, VA, December 8, 2010.

The Future of Nanotechnology Innovation. NNI Innovation Summit, Washington, DC., December 10, 2010.

Biolgocial Interactions of Engineered Nanoparticles: Novel Functions and Nanosafety Issues. PacificChem 2010, Honolulu, HI, December 18, 2010.

High Content and Rapid Throughput Assessment of Nanomaterial's Hazard Potential. PacificChem 2010, Honolulu, HI, December 19, 2010.

Nanomaterial Safety Assessments by Rapid Throughput Approaches. US‐Russia Bilaterial Presidential Commission Nano/EHS Sub‐Working Group Meeting, Moscow, Russia, March 1, 2011.

When is Exposure Not Exposure? Society of Toxicology Annual Meeting, Washington, DC, March 8, 2011.

Human health data needs. NNI Presents US‐EU Bridging NanoEHS Research Efforts. Washington, DC, March 10, 2011.

Roger Nisbet, UC Santa Barbara

Dynamic Energy Budget DEB theory, Scripps Institution of Oceanography, November 2010

Dynamic Energy Budget DEB theory, University of Oldenberg (Germany) Spring 2010

Dynamic Energy Budget DEB theory, Ruder Boskovic Institute, Zagreb (Croatia), Spring 2010


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Suman Pokhrel, University of Bremen

Designing Nanoparticles for Nano‐Bio‐Interaction, ICEIN 2010, University of California, Los Angeles, USA, May 11‐13, 2010.

Tuning the stability and antibacterial properties of ZnO nanoparticles (poster), ICEIN 2010, University of California, Los Angeles, USA, May 11‐13, 2010.

John Priester, UC Santa Barbara

Effects Of CdSe Quantum Dots On Unsaturated Pseudomonas aeruginosa Biofilms (poster) ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Robert Rallo, Universitat Roviri

High Throughput Screening of Nanoparticles Toxicity: Data Mining and Modeling. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Data Mining of High Throughput Screening Toxicity of Engineered Nanoparticles. 2010 AIChE Meeting, November 7‐12, 2010, Salt Lake City, Utah, USA.

Terre Satterfield, University of British Columbia

Exploring the Prehistory of Risk Perceptions: Malleable Perceptions and Upstream Study of the Perceived Risks of Nanotechnology, Annual Meeting of the Society for Risk Analysis, December 8, 2010, Salt Lake City Convention Center, UT

Ambivalence and Nanotechnologies, Annual Meeting of the Society for Risk Analysis, December 8, 2010, Salt Lake City Convention Center, UT

Alia Servin, University of Texas El Paso

Toxicity of TiO2 nanoparticles on cucumber (Cucumis sativus). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Sharona Sokolow, UCLA

UC CEIN Protocols Project: Tools and Guidelines for Development and Validation of Standard Protocols (Poster), ICEIN 2011, Los Angeles, CA, May 11‐12, 2010.

Ponisseril Somasundaran, Columbia University

ACS Akron meeting and ACS San Francisco meeting, before Aug 2010

Sirikarn Surawanvijit, UCLA

Removal of Metal Oxide Nanoparticles from Aqueous Nanoparticle Suspension via pH Adjustment and Coagulation Followed by Membrane Filtration (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Analysis of Membrane Filtration Efficiency in Removal of Metal Oxide Nanoparticles from Aqueous Nanoparticle Suspension in the Presence of Coagulation Pretreatment. Nanotech 2010 Conference, Anaheim, CA, June 2010

Reginald Thio, UC Santa Barbara

Influence of natural organic matter on the adhesion of Au nanoparticles‐coated polystyrene latex beads to mica using “colloid” probe atomic force microscopy. 239th ACS National Meeting, San Francisco, CA, Mar 25, 2010.


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Influence of natural organic matter on the adhesion of Au nanoparticles‐coated polystyrene latex beads to mica using ‘colloid’ probe atomic force microscopy (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Probing the Influence of Solution Chemistry on the Adhesion of Au NPs to mica using colloid probe microscopy, ICEIN 2010: International Conference on the Environmental Implications of Nanotechnology, UCLA, May 12, 2010.

Aggregation and deposition of titanium dioxide (TiO2) nanoparticles as a function of environmental conditions.” 84th ACS Colloid & Surface Science Symposium, Akron, OH, Jun 22, 2010.

Courtney Thomas, UCLA

Stealth and Trojan Horse Nanoparticles. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Magnetically Activated Nanovalve for Drug Delivery.” Second National Cancer Center (Korea)‐UCLA Video Conference on Cancer Research, Los Angeles, CA. December 2010.

Raja Vukanti, UC Santa Barbara

Interactions of engineered nanomaterials with bacterial cell surfaces and effects on growth characteristics (poster). ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Interactions of engineered nanomaterials with bacterial cell surfaces and effects on growth characteristics (poster). American Society for Microbiology 110th Annual General Meeting. 2010. San Diego, CA. May 25, 2010.

Tian Xia, UCLA

Establishment of in vivo zebrafish model for toxicity screening of nanomaterials. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

Kristin Yamada, UCLA

UCLA Sustainable Technology Policy Program (STPP). California Regulation and Prioritization of Nanomaterials (Poster). ICEIN 2010, Los Angeles, CA, May 11 ‐ 12, 2010

UCLA Sustainable Technology Policy Program (STPP). California Regulation and Prioritization of Nanomaterials (Poster). Cell, DNA, and Tissue Damage and Responses Symposium, Riverside, CA, June 4‐5, 2010.

Sharon Walker, UC Riverside

Environmental Implications of Nanotechnology. Ecotoxicology Graduate Program Seminar, University of California, Riverside, October 20, 2010

Jeffrey Zink, UCLA

SPIE meeting in San Francisco

the National ACS meeting in SF

DTRA meeting in Washington

DTRA meeting DC in Orlando, FL Dongxu Zhou, UC Santa Barbara

Mechanisms Responsible for the Initial Aggregation of Metal Oxide NanoParticles in Water. ICEIN 2010. Los Angeles, California, May 11‐12, 2010.

poster presentation at the 2010 ACS Spring meeting in San Francisco


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poster at 2010 Goldschmidt conference in Knoxville, TN Outreach Activities by UC CEIN Members Collaborations with International Researchers

Chinese Academy of Sciences

University of Tsukuba, Japan

University of Kyoto, Japan

National Institute of Chemical Physics and Biophysics Akadeema, Estonia

CINVESTAV, Mexico

Instituto Nacional de Salud Publico (INSP), Mexico Collaborations with Industry

Held initial discussions with and hosted visit to UCLA by Seth Coe‐Sullivan, QD Vision to explore potential collaboration on environmental fate and transport and safe handling of Quantum Dots. February, 2011.

Visit and discussion with Byron Cheatham of Cytoviva. Visits to both UCLA and UCSB to discuss development of imagining technology for use in nanomaterial studies conducted by the Center.

Formed a partnership with the Center for High Rate Nanomanufacturing at Northeastern University (Ahmed Busnaina, Director) to explore partnerships with Industry geared towards safety testing of nanomaterials using alternative testing strategies. A first collaborative workshop between academia, industry, and regulatory agencies is planned for October 2011, with a focus on Carbon Nanotubes.

Legislative/Policy Activities

CEIN members Andre Nel, Barbara Harthorn, Hilary Godwin, and C. Jeffrey Brinker played key roles in World Technology Evaluation Center Nano 2 project, contributing to 4 chapters in the recently published review of the future of Nanotechnology research in the next 10 Years entitled: "Nanotechology Long‐Term Impacts and Research Directions: 2000 ‐ 2020."

UC CEIN Director Andre Nel chosen to serve on the Bilaterial Presidential Commission for US/Russia Cooperation.

Educational Mentoring Bradley Cardinale, UC Santa Barbara

Two high‐school interns (Chelse Tsai‐Simek and Aaron Juarez) completed CEIN research projects in the lab during the summer of 2010 as part of UCSB’s High School Mentorship Program. We are presently finishing a manuscript for submission of Chelse’s research, which examined trophic transfer of nano‐TiO2 from algae to snails.

Patricia Holden, UC Santa Barbara

Mentored (educated and trained) two masters student interns (6 months each) from the Bren School Master of Environmental Science & Management (MESM) program: Adeyemi Adeleye and Shivira Tomar. Each was trained in nanotoxicology basics, basics in PHB production and importance, basic lab methods, and was trained to perform specific tasks needed for this research.

Mentored (educated and trained with Joshua Schimel) Vivian Chang, a high school student intern (for 6 weeks in the Summer ‘10) from the UCSB Summer Research Mentorship Program.


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The intern was trained in nanotoxicology and soil physics basics, basic lab methods, and was trained to perform specific tasks needed for this research.

Arturo Keller, UC Santa Barbara

Mentored 3 undergraduates for CEIN work – Jon Conway, Gabriel Rubio and Ingmar Prokop on dispersing Ag and TiO2 nanoparticles in aqueous media, measuring their sizes and zeta potentials with Zetasizer, using the TLC plates to determine Ag nanoparticle mobility in aqueous media and utilizing the UV‐VIS spectrophotometer to measure Ag and TiO2 nanoparticle concentrations in water.

Roger Nisbet, UC Santa Barbara

Kaysha Nelson has initiated an undergraduate honors project at UCSB that examines how food‐web complexity influence bioaccumulation of nano‐TiO2, under the mentorship of Konrad Kulacki and Roger Nisbet (IRG 2).

Sharon Walker, UC Riverside

Undergraduate research mentoring (James Kim)


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13. Shared and other Experimental Facilities UCLA Facilities The UC CEIN is housed in the California NanoSystems Institute (CNSI) building, centrally located on the UCLA campus. Administrative office space for CEIN support staff and access to CNSI meeting rooms, conference rooms, state of the art media facilities, and meeting planning assistance are provided within CNSI. Additionally, over 1000 square feet of shared laboratory bench space has been allocated to CEIN researchers. The CEIN recently installed a Quadrasorp SI to analyze surface area and pore size of CEIN nanomaterials and is in process of installing a Perkin‐Elmer AAnlyast Graphite Spectrometer. This equipment follows the recently installed Wyatt DynaPro Plate Reader Dynamic Light Scattering instrument, a Brookhaven Zeta Potential analyzer, and an Elisa Plate reader for use by CEIN researcher in characterization and high content screening studies. Bench space has also been outfitted to accommodate approximately 10 working bays. Finally, the Data Management activities have the Center have been assigned private space in the CNSI Data Center, providing workstations and a server room to serve as the hub of the UC CEIN data management and modeling activities as described in IRG 6. Molecular Screening Shared Resource (MSSR): Established in 2003, the MSSR provides HTS technology. Since 2005, it has been directed by Kenneth Bradley, who reports to an Advisory Board with members from UCLA’s CNSI, Chemistry, Biology, Medicine, and other departments. Located in the UCLA CNSI, the MSSR occupies 1085 square foot of laboratory space. The MSSR contains two fully integrated systems: (i) Automated liquid handling, multiple plate reading, plate filling and washing, deshielding, and delidding, and online incubators for cell‐based assays using a Beckman/Sagian system equipped with an Orca robotic arm that delivers plates to individual work stations; Beckman Biomek FX liquid handling robot (96‐well pipetting, 96‐ or 384‐pin transfer); Perkin–Elmer Victor3(V) plate reader (96–1536 well plates in luminescence, fluorescence, fluorescence polarization, time‐resolved fluorescence, UV–Vis absorbance modes); Molecular Devices FlexStation II plate reader equipped with an integrated pipetter and general fluorescence and luminescence plate applications in 96‐ or 384‐well format; Cytomat 6001 incubator: CO2 incubator; Multidrop 384: manifold liquid dispensing into 96‐ or 384‐well plates; ELx 405 plate washer: well washing, aspiration, dispensing. Current capacity of cell‐based assay is ca. 105 wells (conditions)/day. Multiple plate readers allow fluorescence, FRET, BRET, time‐resolved fluorescence, fluorescence polarization, luminescence, and UV–Vis absorption assays. (ii) A second Beckman/Sagian Core system for HCS using automated microscopy with an Orca arm; Molecular Devices ImageXpress (micro) automated fluorescence microscope and a Cytomat 6001 incubator. Equipment at MSSR available for off‐line use: Genetix Q‐bot colony‐picking robot: maintain and re‐order clone collections; Precision 2000: automated pipetting and manifold dispensing of BSL‐2 agents; 6‐ft Class II biosafety cabinet, tabletop centrifuge, –80 °C freezers, 96‐well thermal cycler, CO2 incubators. MSSR screening capabilities include two genome‐wide knockout libraries of S. cerevisiae yeast and genome‐wide small interfering RNA libraries for mouse and human, providing functional genomic capabilities for identifying cellular pathways governing responses to nanomaterials. The following research facilities on the UCLA campus are available to the CEIN on a recharge basis: CNSI Core Facilities including an Advanced Light Microscopy/Spectroscopy, X‐ray Diffraction, and Imaging Lab, Electron Imaging Center for NanoMachines, Integrated NanoMaterials Lab, Integrated Systems Nanofabrication Clean Room, Macro‐Scale Imaging, Molecular Screening Shared Resource, Nano & Pico Characterization. Water Quality Research Laboratory includes NF/RO membrane simulators for desalination and wastewater reclamation studies, MF/UF membrane simulators with rapid permeate back‐flushing, integrated membrane filtration/mixed reactors for hybrid membrane process studies, flow through electrochemical reactor for electrodialysis and electro‐oxidation studies.


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Water Quality Instrumentation Laboratory includes GCs with FID, ECD, MS, and TCD detectors, HPLC, ion and hydrophobic chromatography, flame atomic absorption spectrophotometer for trace metal analyses, TOC, UV–Vis, and fluorospectrophotometer for organic characterization and analyses and instrumentation for pH, conductivity/TDS, ions, turbidity, color, particle size, and solids analyses. Water Technology Research Center includes atomic force microscopy (AFM), IR spectroscopy (FTIR, ATR‐IR), X‐ray photoelectron spectroscopy. Nanoelectronics Research Facility includes scanning electron microscopy (SEM) with energy‐dispersive analysis of X‐rays; transmission electron microscopy; surface profilometers and ellipsometers. Molecular Instrumentation Center includes SEM, differential scanning calorimetry, thermogravimetric analysis, magnetic resonance imaging, X‐ray diffraction, mass spectrometry for proteomics and biochemistry instrumentation, ICP‐AES for elemental analysis and speciation. UCLA’s Environmental Nanotechnology Research Laboratory includes a programmable oven, furnace, and microwave systems for NM synthesis, bench‐top micro‐centrifuge and stirred filtration cells for NM isolation, BET analyzer for powder surface area and pore size analyses, equipment for polymer phase inversion, interfacial polymerization, and solution casting. Nano‐Bio Interfacial Forces Laboratory includes a contact angle goniometer for powder/substrate wetting and surface energy analyses; particle micro‐electrophoresis system for particle electrophoretic mobilities (zeta potentials); dynamic and static light scattering for evaluating particle sizes and polymer molecular weights; upright optical and epi‐fluorescence microscope; and AFM integrated with inverted optical microscopy. UC Santa Barbara Facilities Three clusters of laboratories are available to CEIN researchers: (1) CNSI‐UCSB provides access on a recharge basis: The Microscopy and Microanalysis Facility includes three transmission electron microscopes (FEI Titan FEG and two FEI Tecnai G2 Sphera), three SEMs (FEI XL40 Sirion FEG, FEI XL30 Sirion, FEI Inspect S), five scanning probe STM/AFM microscopes (Digital Multi‐mode Nanoscope, Digital Dimension 3000, Digital Dimension 3100, Asylum MFP‐3D SL, Asylum MFP‐3D Bio), a secondary ion mass spectrometer (Physical Electronics 6650 Quadrupole), X‐ray Photoelectron Spectroscopy Kratos Axis Ultra System, Focused Ion Beam System (Model DB235 Dual Beam). The Spectroscopy Facility has seven state‐of‐the‐art spectrometers (Nicolet Magna 850 IR/Raman, Varian Cary Eclipse Fluorimeter, Bruker DPX200 SB NMR for solutions, DSX300 WB NMR for solids, DMX500 SB NMR for solutions, Bruker IPSO500 WB NMR for solids, Bruker EMX Plus EPR spectrometer). Stucky’s laboratory (3000 sf) contains: Malvern Nano‐sizer, Invitrogen Xcell SureLock Mini‐Cell Gel Electrophoresis, Nikon Eclipse ME600 Microscope with CCD camera, Arbin Instruments, MSTA+ & EG&G Princeton Applied Research potentiostat/galvanostat, Netzsche STA 409C thermogravimetric analyzer, Tempress hydrothermal system, Brinkman Tuttnauer autoclave, humidity‐environment‐controlled reaction chamber, vacuum oven, 1‐gallon Parr autoclave reactor, three vacuum‐atmosphere boxes, Labconco Free Zone 4.5‐L benchtop freeze dry system, IEC Multi‐RF high‐performance centrifuge, OLIS Cary 14 UV–Vis–near‐IR spectrophotometer, StellarNet UV–Vis spectrophotometer with a photodiode detector. (2) Bren School of Environmental Science and Management. The School Infrastructure Lab (2350 sf) includes a Shimadzu HPLC with fluorescence and diode array detectors, Shimadzu GC/FID, Beckman scintillation counter, total‐carbon analyzer, –80 °C Revco freezer, high‐speed refrigerated Sorvall centrifuge, two static incubators for cultivation at 37 and 41 °C, refrigerator, water baths, spectrophotometers, hybridization oven, UV crosslinker, Nanopure water system, autoclave, icemaker, laboratory microwave, two multi‐user walk‐in 4 ºC rooms for sample storage and two walk‐in freezers, and two variable‐temperature rooms for experimental work. Use of this central facility is available at no cost to the project. Holden’s laboratory (930 sf) includes: HP 6890 GC/MS with autosampler; Baker


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biological control cabinet; Sorvall microcentrifuge; New Brunswick shaker/incubator; analytical balances; Nikon E‐800 epifluorescent microscope equipped with a CCD camera and NIS‐Elements acquisition and analysis software; BioTek Synergy2 microplate shaker/incubator/reader with UV/Vis/TRF detectors; PCR thermal cycler and other equipment related to electrophoresis, PCR product quantification, and analyzing terminal labeled restriction fragment length polymorphisms. Holden’s laboratory houses the Micro‐Environmental Imaging and Analysis Facility (MEIAF), an environmental SEM with a cryo‐stage for imaging frozen materials and an X‐ray detector for elemental analysis (300 sf). The MEIAF is available to the public on a recharge basis. Keller’s laboratory (940 sf) includes: Varian Saturn 2100T GC/MS with autosampler; Nikon Optiphot‐M epi‐fluorescent microscope with CCD camera; Thermo Cahn Radian 315 dynamic contact angle analyzer; Brookfield viscometer; ozone generator; UV reactor; column transport pumps and controllers; silicone micromodels. The School has a Videoconferencing Facility (Bren Hall 1424, 750 sf) for telecommunication (e.g., graduate training courses, seminars, REC meetings) related to the project, with capacity for 50 people. It supports h242 video conferencing or ISDN over IP connections at 100 Mbps. There are two data projectors and corresponding screens that can display input from a remote video connection or from local inputs including a dedicated computer or portable DVD/VHS video camera and a document camera. (3) Department of Ecology, Evolution, and Marine Biology. Laboratories from three EEMB faculty will be used for the proposed work. Schimel’s laboratory includes: two Finnegan MAT Delta Plus MS systems equipped with elemental analyzer, gas bench, pyrolysis, and GC inlet systems (available through MSI analytical lab); two multichannel Lachat autoanalyzers for dissolved nutrients; C/N analyzer for solid samples; Shimadzu GC 14 for simultaneous CO2, CH4, and N2O analyses; microtiter plate reader (UV/Vis) for enzyme and chemical assays. Cardinale’s laboratory (1200 sf) is fully equipped for work with aquatic algae and invertebrates; it includes: two environmentally controlled walk‐in chambers for the culture of organisms; a “clean lab” equipped with a Millipore water purification system for nutrient analyses and water chemistry; an 800‐sf state‐of‐the‐art freshwater flume facility (temperature‐controlled facility with 120 recirculating stream channels); two Olympus stereomicroscopes for invertebrate work; Barnstead–Themolyne spectrophotometer; Turner fluorometer; two YSI Model 556 oxygen probes; Sontek Flowtracker Acoustic Doppler Velocimeter for field work; Li‐Cor LI‐192 underwater quantum sensor with LI‐1400 datalogger. Nisbet’s laboratory has high‐end PCs for DEB modeling; it has substantial computing requirements and requires Linux and Windows applications. Additional access to a high‐performance computing multi‐node facility at UCSB is available on a recharge basis. Lawrence Berkeley National Laboratory Facilities Molecular Foundry. Molecular Foundry users apply to use the facilities and are trained by LBNL staff to conduct their studies. The Inorganic Nanostructures Facility will provide CEIN users with the instrumentation to synthesize and characterize nanocrystals, nanotubes, and nanowires, as well as their expertise and training on manufacturing processes. The following equipment is available free of charge: Automated nanocrystal synthesizer robot; Bruker AXS D8 Discover GADDS XRD diffractometer system; Thomas Swann 3x2 CCS MOCVD for nitride films and nanowires; Thomas Swann 3x2 CCS MOCVD for III‐V and other semi‐conducting materials; Yobin Ivon Fluorolog 3 spectrofluorimeter with PL life‐time capability; Agilent Precision semiconductor parameter analyzer; Malvern Zetasizer ZS; Shimadzu UV–near IR spectrophotometer; Rucker and Kolls probe station; low‐temperature, inert‐atmosphere probe station; custom‐built robotic combinatorial synthesizers; Beckman NXp HTS robot; total‐internal‐reflection microscopy system equipped with Olympus IX‐81w/Andor EMCCD camera; Amersham Biosciences Akta FPLC; Agilent 1100 series (ion trap) LC‐MC‐MC mass spectrometer; Varian analytical, semi‐prep, and prep HPLCs; CEM Liberty microwave peptide synthesizer; Biotage SP1 flash chromatography system; ACT Apex 396 peptide synthesizer; Beckman Optima ultracentrifuge; Real‐time


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PCR 7000 sequence detection system; New Brunswick BioFlo 310 fermentor; Molecular Devices absorbance and fluorescence plate readers; Jobin Yvon FluoroMax flourimeter; and cell culture incubators and biosafety cabinets. Columbia University Facilities The Industry/University Cooperative Research Center (I/UCRC) for advanced studies on novel surfactants has shared resources in the MRSEC and Chemistry Departments for work on this project: Hitachi 4700 SEM; JEOL SEM and TEM; Inel X‐ray diffractometer; Bruker NMR spectrometer; Raman spectroscope; ellipsometer. Somasundaran’s laboratory includes: Digital Instruments AFM; PenKem 3.0+ Zeta meters; Perkin–Elmer Spectrum100 FTIR spectrophotometer; Horiba Jobin Yvon Fluorolog fluorescence spectrophotometer (steady state); Horiba Jobin Yvon IBH5000F fluorescence spectrophotometer (time‐resolved); Quantachrome Instruments Quantasorb surface area analyzer; Bruker EMX EPR spectroscope; Perkin–Elmer Plasma 400 ICP spectrophotometer; Kruss K12 surface and interfacial tensiometers; NIMA Tech DST9005 dynamic surface tension analyzer; Nikon optical microscope; Beckman–Coulter Optima XL‐1 analytical ultracentrifuge; SORVALL RC‐5B bench‐scale and temperature‐controlled centrifuge. University of Bremen Facilities Foundation Institute for Materials Science. The IWT Foundation Institute of Material Science has all major material characterization equipment available: X‐ray diffraction (with extended Rietveld analysis); TEM and SEM; surface adsorption analysis (adsorption isotherms). Recharge‐based access to thermogravimetric analysis and zeta‐potential instrumentation is also available. Mädler’s laboratory has state‐of‐the‐art flame spray pyrolysis reactors for the synthesis of various metal oxide‐based NMs, including their functionalization with noble metals. UC Riverside Facilities Center for Nanoscale Science and Engineering. A 1900‐sf laboratory space with environmental controls necessary to provide Class 1000 and Class 100 clean areas. CNSE has four staff members: a Facility Manager, a dedicated operator for the e‐beam and FIB instruments, and maintenance and process technicians. In addition to the nanofabrication center, the following major resources are available on a recharge basis: NMR spectroscopy; mass spectrometry; small‐molecule X‐ray crystallography (SMXC); optical spectroscopy; fluorescence‐activated cell scanner (to analyze cell morphology, cell surface proteins, and cell cycle‐related processes); high‐precision CNC lathe; mill and Sinker‐ and Wire‐EDM; electron beam techniques; laser confocal microscopy. Walker‘s laboratory at UCR is equipped with an inverted Olympus IX70 microscope (phase contrast or fluorescent mode), used to image bacterial cells or particle attachment to test surfaces within a parallel plate flow cell or a radial stagnation point flow cell. Image analysis software allows quantification of the kinetics of cell–particle attachment to the test surfaces. Nanyang Technological University (NTU) Facilities. Boey’s laboratory at NTU has the following equipment for characterizing NMs for the proposed work: dynamic light scattering; zeta potential analyzer; FE‐SEM; HR‐TEM; XPS; MALDI‐TOF MS; ASAP‐BET; FTIR spectrometer; a range of XRDs. UC Davis Facilities Bodega Marine Laboratory (BML). BML has an outstanding flow‐through seawater system, a sophisticated computer‐controlled 600,000‐gallon/day system providing seawater to 16 wet lab areas. A Seawater Monitoring and Control Network provides automated and centralized control of temperature in 10 labs and salinity in two. Photoperiod control is also available in several areas and natural sunlight


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in two outdoor laboratories. Other functional spaces include 49 dry laboratories, three classrooms, and one auditorium as well as a library, computer lab, and two conference rooms. BML is also equipped with a Horiba JY Ultima 2C ICP‐OES (optical emissions spectrometer); New Wave laser ablation system; two semi‐automated quantitative fluorescence imaging systems (Olympus Fluoview 500 scanning laser confocal; Metamorph/Metafluor imaging system with cooled high‐speed CCD); Tecan fluorescent plate reader. Cherr’s laboratory houses the BML’s Fluorescence Imaging Facility, which includes a Photon Technology spectrofluorometer with ratiometric and ion quantitation software; high‐speed fluorescence video imaging system; three epifluorescence microscopes; UVP Epichem II fluorescence/chemiluminescence gel documentation system; Tecan Genios time‐resolved fluorescence/ and luminescence/absorbance plate reader; confocal scanning laser microscope; Expert Vision System software. This facility is equipped to analyze motion of microscopic samples as well as larger macro samples (e.g., adult fish and crustaceans). University of Texas, El Paso Facilities Gardea‐Torresdey’s laboratory has available the following major equipment for this project: 3100 Perkin–Elmer flame atomic absorption spectrometer; 4100 ZL Perkin–Elmer Zeeman graphite furnace atomic absorption spectrometer; 4300 DV Perkin–Elmer ICP OES; Perkin–Elmer Elan DRC IIe Laser ablation/HPLC/ICP‐MS; EG&G Model 394 electrochemical trace analyzer; Hewlett–Packard 5890 GC; Hewlett–Packard 5972 GC/MS; Perkin–Elmer Spectrum 100 FTIR spectrometer coupled to a Perkin–Elmer Spectrum spotlight 300 FTIR microscope. Additional shared resources: Bruker 250‐MHz NMR spectrometer; Bruker 300‐MHz multi‐nuclei NMR spectrometer; Electroscan 2020 environmental SEM; Kevex omicron X‐ray microfluorescence spectrometer; Hitachi S‐4800‐II SEM with EBSD; EDAX/TSL X‐ray analyzer and electron backscatter diffraction imaging equipment; Zyvex Nanomanipulator and Nanoprobe; Hitachi H‐8000 TEM. The XAS studies planned for this project will be performed at Stanford Synchrotron Radiation Laboratories (SSRL), Stanford, CA, where Gardea‐Torresdey has received beam time for performing X‐ray absorption spectroscopic studies for the duration of this project. Sandia National Lab Facilities Brinker's Biocharacterization laboratory includes a facility for the integration of biological organisms/components with engineered platforms. The lab is capable of handling Level 2 biological organisms and the isolation and analysis of DNA, RNA, and proteins. Various methods are used to incorporate biological organisms/components onto engineered platforms, such as vesicle fusion, multiple tethering schemes, and plugged flow packing. Other capabilities include: ellipsometry for film characterization; electrochemistry; a PCR instrument for DNA amplification; a laser connected to an inverted microscope for fluorophore interrogation; and a hyperspectral microarray scanner for microarray analysis. A Biosafety Level 1 laboratory is in operation at the AML with access to BSL2 status facilities at Sandia. The AML facility contains standard microbiological and biochemical equipment and supplies for handling the microorganisms and cell lines proposed for use on this project: Class II flow bench; standard and CO2 incubators; cryo‐storage; freezers and refrigerators; autoclave; and a fluorescence microscope. In Spring 2008, Sandia will install an Asylum Research MFP‐3D‐BioAFM integrated with a Nikon TE2000‐U inverted fluorescence microscope, which combines molecular resolution imaging and picoNewton force measurements on an inverted optical microscope to allow: in situ imaging of the surfaces of living cells upon exposure to NMs; measurement of adhesive forces of proteins/NMs on cell surfaces; single‐molecule force spectroscopy of single NPs; and nanolithography and manipulation of samples on the nanometer and picoNewton scale.


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Northwestern University Facilities The Hersam Laboratory encompasses 3000 square feet distributed among five rooms in Cook Hall at Northwestern University. This space houses five fume hoods and the following major pieces of instrumentation:

2 Thermomicroscopes CP Research Atomic Force Microscopes (AFMs): These instruments characterize mechanical (force‐distance spectroscopy) and electronic (electric force microscopy and scanning potentiometry) properties of materials at the nanometer scale in ambient, controlled atmosphere, and liquid environments. We have also employed these AFMs for nanolithography (e.g., field‐induced oxidation and liquid phase nanolithography).

2 Room Temperature Ultra‐high Vacuum (UHV) Scanning Tunneling Microscopes (STMs): These home‐built multi‐chamber systems are used to prepare pristine surfaces, which are then characterized at the atomic‐scale with STM and scanning tunneling spectroscopy. Although predominantly used at room temperature, these systems can also scan samples at elevated temperatures up to 700 K. Feedback controlled lithography has also been implemented to isolate and pattern individual molecules on surfaces in atomically precise geometries. The UHV chambers (base pressure ~ 2×10‐11 Torr) are directly interfaced to a controlled atmosphere glove box (oxygen and water concentrations < 1 ppm) to enable combined UHV and wet chemical processing with minimal contamination.

1 Cryogenic Variable Temperature UHV STM: In addition to all of the capabilities of the room temperature UHV STMs, this system also controls the temperature of the sample and the microscope between 10 K and 400 K. Consequently, this microscope is ideal for cryogenic studies and high resolution scanning tunneling spectroscopy.

1 Nanoelectronic Charge Transport Measurement Apparatus: This system enables electrical characterization of nanoscale devices and sensors. The apparatus includes a wafer prober, hall measurement apparatus, high sensitivity source‐measure unit, spectrum analyzer, current preamplifier, lock‐in amplifier, and 4‐channel digital oscilloscope.

3 Density Gradient Ultracentrifugation (DGU) Apparatuses: These systems are used to sort carbon nanotube and graphene samples by their physical and electronic structure. Each apparatus includes a horn ultrasonicator, a Beckman Coulter Optima L‐90 K Preparative Ultracentrifuge, and a BioComp Piston Gradient Fractionator. In addition, a Cary 5000 UV/Vis/NIR spectrophotometer is available for post‐DGU sample characterization.


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14. Personnel Management and Organization Strategy The UC CEIN strategy is to maintain a strong organizational infrastructure that supports and integrates our research, technology development, educational and diversity efforts, internal and external stakeholders, as well as facilitating seamless communication among all these communities. To this end our organizational structure allows for selection, prioritization, distribution, and management of resources within a multi‐institutional center structure. Leadership Andre Nel (UCLA) serves as the Center Director and Principal Investigator. As Director, Dr. Nel is responsible for the integration of the Center’s overall research, education and outreach activities. Arturo Keller (UCSB) is the Associate Director, responsible for coordinating the research integration, seminars, student training, and outreach activities at UC Santa Barbara to provide seamless integration with the activities at UCLA. Focused leadership for the education and outreach components of the Center is provided by Hilary Godwin (UCLA). This faculty management team provides complimentary expertise and strategic leadership to ensure the Center’s vision and mission. Integrated Research Groups (IRGs). CEIN research is organized into seven research groups, each under the leadership of a CEIN faculty member. Each IRG is composed of several faculty, postdoctoral researchers, research staff, and graduate students. Key to the success of the CEIN is the integration of research within and across IRGs. IRG leaders are responsible for setting priorities, allocating resources, and tracking progress towards achievement of IRG goals. Frequent formal communication between IRG leaders is key to ensuring that progress is made across all groups, and the findings of one IRG are rapidly disseminated other IRGs. Projects submit quarterly progress updates to their IRG leader, the results of which are shared and discussed by the CEIN Executive Committee. Executive Committee The Executive Committee is composed of the Director, Associate Director, Education/Outreach Director, Co‐PIs, IRG leaders, and the Center Chief Administrative Officer. The Executive Committee meets at least once per month and is responsible for assisting the Director with integration and coordination of research and education, overall resource allocation, and outreach to the scientific, industrial, and policy community. Each quarter, the Executive Committee reviews long‐term directions of the Center and possible strategic redirections. Prior to any Research Reviews, Site Visits, and External Science Advisory Committee meetings the EC focuses on strategic planning. Research progress for all projects is reviewed on an ongoing basis, with projects submitting Quarterly progress updates. Allocation of Center resources is based on the following metrics: (i) contribution of the proposed work to the CEIN’s core goals; (ii) productivity, publication, and product delivery record; (iii) novelty; (iv) integration and cooperation with other funded CEIN projects; (v) availability of resources and facilities to carry out proposed projects; and (vi) timely delivery of tangible results. Approximately 5% of the total research budget is designated for new and exploratory integrated research seed funding. Proposals for seed funding are reviewed by the EC on an annual basis. Each March, the Executive Committee meets for a day long research retreat. The retreat focuses on the review of overall Center priorities and is a forum for discussing and establishing key short and long term goals for the Center, with particular focus on strengthening integration across all IRGs.


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External Science Advisory Committee The CEIN has convened an 11‐member External Science Advisory Committee (ESAC) comprised of scientists, technologists, industry members, and policy and education specialists. The ESAC advises the Center’s Executive Committee with respect to CEIN strategic directions and management policies. The ESAC provides feedback on the focus and direction of CEIN research, progress made toward achieving Center goals, and illuminating new research and educational opportunities. The diversity of this group provides a comprehensive perspective on the major advances in nanotechnology and key issues with regards to potential environmental implications. After two initial meetings with Center leadership via videoconferencing technology, the first in‐person meeting of the ESAC took place in May 2010 on the UCLA campus. This daylong meeting involved in depth discussions about the CEIN approach and organization of Center research and integration. The ESAC provided valuable advice on the strengths of the Center in advance of the June 2010 NSF Site Visit. The next ESAC videoconference will be held in May 2011, followed by an in‐person visit in Fall 2011. This next round of ESAC meetings will seek advice as the Center prepares for renewal. Student‐Postdoctoral Advisory Committee A Student‐Postdoctoral Advisory Committee (SPAC) continues to be active within the CEIN. The committee includes graduate student and postdoctoral scholar representatives from each of the IRGs. The SPAC provides ongoing input into the development of the CEIN education program (including development of undergraduate mentoring opportunities), development of full‐day annual leadership workshops (held this year in May 2010 and March 2011), and formulation of goals for future Center workshops and seminar series. With input from the SPAC, the Education/Outreach Director and Coordinator have developing an assessment document reviewing the educational and training achievements of Center trainees, results of which are discussed with the SPAC. Administrative Support An administrative staff has been compiled at UCLA to support streamlined operations of the Center. David Avery serves as the Chief Administrative Officer of the CEIN. The CAO assists the Director by overseeing the general administration, cooperation, communication, planning, financial implementation, goals setting, and development of Center activities. The CAO is supported by the following dedicated staff:

o Financial/Budget Coordinator – responsible for financial management and reporting systems across partner institutions

o Administrative Assistant – provides general support for all Center activities including meeting coordination

o Education/Outreach Coordinator – under joint supervision of the CAO and Education/Outreach Director, organizes the training, communication, diversity, and evaluation components of the program.

To assist in the administrative coordination of the UC Santa Barbara activities, a half time administrative support staff position has been allocated to UCSB.


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Organization Chart

IRG 1 Leader

IRG 2 Leader

IRG3 Leader

IRG4 Leader

CEIN Exec utive Committee (CEC)

CEIN Director

External ScienceAdvisory Committee

( ESAC )

CEIN Associate Director

Student

Advisory Committee

SPAC

IRG5 Leader

Modeling of NP Environmental Distribution &

Toxicity

IRG 6 Leader

Combinatorial Nanoparticle

Libraries

NMs Interactions ( Molecular ,

Cellular , Organ & Systemic

Levels )

Organismal & Community Toxicology

Nanoparticles Fate &

Transport

High Throughput Screening (Data Mining, QSRs)

IRG7 Leader

Risk Perception

Education / Outreach ( EO )

Director

Vice Chancellor for

Research

Education , Outreach & Human

Resource Objectives

Chief Admin

Officer

Financial/Budget

Coordinator

Administrative Assistant

EO Coordinator /

Assistant

Administrative Assistant

Data/IT Coordinator

Postdoc

Changes in Personnel As part of the annual review of the Center research priorities, IRG 5 Leader Kenneth Bradley requested that IRG 5 be reorganized to reflect his intellectual interest for the upcoming year. While IRG5 has made an important contribution under the leadership of Dr. Bradley, he has requested that his major interest pertains to the science of high throughput screening a rather than the technicality and complexity of nanotechnology in which he has no official training. He is very interested, however, is continuing to play an active role as the faculty director of the Molecular Shared Screening Resource (MSSR) while the specifics of nanomaterials screening is handled under an IRG leader that is schooled in nanotechnology. Thus, under the proposed reorganization, Dr. Bradley will serve as the Technical Director of the Molecular Shared Screening Resource, which will assume a core function role within the Center. Dr. Robert Damoiseaux will continue to assist Dr. Bradley by providing technical consultation and assistance in the planning of high throughput experiments, including the translation of assays to HT capabilities. Scientific leadership of IRG 5 will be transferred to Dr. Andre Nel, who is proficient in mammalian high throughput screening and will be assisted by doctors Hillary Godwin and Dr. Ken Bradley in continuation and expanding bacterial high throughput screening. Selection of Dr. Nel as IRG 5 leader has been discussed and agreed upon by the Executive Committee.


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At the March 2010 Executive Committee retreat, the Center Executive Committee agreed to prioritize the introduction of variations of single walled carbon nanotubes into our nanomaterial libraries, including a highly purified form of nanotube that can be used compared against commercially obtained carbon nanotubes. Dr. Mark Hersam from Northwestern University was identified by the Executive Committee as a researcher with the technical capabilities to produce and characterize materials to the specifications needed to introduce into our biological and environmental studies. His research agenda is complementary to the research of the CEIN. Funds from this project were allocated from our IRG 1 nanomaterial acquisition budget and Dr. Hersam's participation allows us to expand our research beyond commercially available carbon nanotubes. Dr. Hersam's biographical information follows in Section 16.

Table 4a: NSEC Personnel - All, irrespective of Citizenship - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Personnel Type Total

Gender Race Data


%NSEC

DollarsMale Female AI/AN NH/PI B/AA W A



W/A

NotProvided

Leadership, Administration/Management

Subtotal 15 7 8 0 0 0 13 2 0 0 0 1 0 100%

Directors1 2 2 0 0 0 0 2 0 0 0 0 1 0 100%

Thrust Leaders1 7 4 3 0 0 0 7 0 0 0 0 0 0 100%

AdministrativeDirector andSupport Staff

6 1 5 0 0 0 4 2 0 0 0 0 0 -

Research

Subtotal 153 92 60 0 0 0 97 52 0 1 2 12 64 94%

SeniorFaculty1 27 24 3 0 0 0 22 5 0 0 0 4 11 74%

Junior Faculty1 1 1 0 0 0 0 1 0 0 0 0 0 1 100%

Research Staff 20 12 8 0 0 0 13 5 0 0 2 1 8 -

VisitingFaculty1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

IndustryResearchers 0 0 0 0 0 0 0 0 0 0 0 0 0 -

Post Docs1 35 22 13 0 0 0 17 18 0 0 0 1 27 100%

DoctoralStudents1 43 21 22 0 0 0 26 17 0 0 0 3 15 98%

MastersStudents1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

UndergraduateStudents (non

REU)126 12 14 0 0 0 18 7 0 1 0 3 2 100%

High SchoolStudents 1 0 0 0 0 0 0 0 0 0 0 0 0 -

Curriculum Development and Outreach

Subtotal 20 8 12 0 0 1 9 6 1 0 3 1 1 61%

SeniorFaculty1 2 2 0 0 0 0 2 0 0 0 0 0 0 50%

Junior Faculty1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

Research Staff 2 1 1 0 0 0 2 0 0 0 0 0 0 -



Post Docs1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%




REU)14 1 3 0 0 0 1 1 1 0 1 0 0 0%


REU Students

Subtotal 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

REU studentsparticipating in NSEC

Research10 0 0 0 0 0 0 0 0 0 0 0 0 0%

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NSEC FundedREU Students1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

Pre-college(K-12)

Subtotal 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

Students 0 0 0 0 0 0 0 0 0 0 0 0 0 -

Teachers (RET) 0 0 0 0 0 0 0 0 0 0 0 0 0 -

Teachers(non-RET) 0 0 0 0 0 0 0 0 0 0 0 0 0 -

Total1 188 107 80 0 0 1 119 60 1 1 5 14 65 77%

1 The percentage of people in the personnel category receiving at least some salary or stipend support from NSF NSEC Program must be provided in the far right column, "%NSEC Dollars." Details are described in the Instructions section for this table.

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Table 4b: NSEC Personnel - US Citizens and Permanent Residents - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Personnel Type Total

Gender Race Data


%NSEC

DollarsMale Female AI/AN NH/PI B/AA W A



W/A

NotProvided

Leadership, Administration/Management

Subtotal 15 7 8 0 0 0 13 2 0 0 0 1 0 100%

Directors1 2 2 0 0 0 0 2 0 0 0 0 1 0 100%

Thrust Leaders1 7 4 3 0 0 0 7 0 0 0 0 0 0 100%

AdministrativeDirector andSupport Staff

6 1 5 0 0 0 4 2 0 0 0 0 0 -

Research

Subtotal 89 51 37 0 0 0 70 15 0 1 2 9 0 99%

SeniorFaculty1 16 14 2 0 0 0 14 2 0 0 0 1 0 94%

Junior Faculty1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

Research Staff 12 7 5 0 0 0 10 0 0 0 2 1 0 -



Post Docs1 8 4 4 0 0 0 8 0 0 0 0 1 0 100%




REU)124 11 13 0 0 0 17 6 0 1 0 3 0 100%


Curriculum Development and Outreach

Subtotal 19 7 12 0 0 0 9 6 1 0 3 1 0 59%

SeniorFaculty1 2 2 0 0 0 0 2 0 0 0 0 0 0 50%

Junior Faculty1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%

Research Staff 2 1 1 0 0 0 2 0 0 0 0 0 0 -



Post Docs1 0 0 0 0 0 0 0 0 0 0 0 0 0 0%




REU)14 1 3 0 0 0 1 1 1 0 1 0 0 0%


Total1 123 65 57 0 0 0 92 23 1 1 5 11 0 77%

1 The percentage of people in the personnel category receiving at least some salary or stipend support from NSF NSEC Program must be provided in the far right column, "%NSEC Dollars." Details are described in the Instructions section for this table.

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UC CEIN Publications

15. Publications and Patents Year 3 ‐ April 1, 2010 ‐ March 31, 2011 Primary Project Publications ‐ Journals 1. Beaudrie, C.E.H., Kandlikar, M., Horses for Courses: Risk Information and Decision Making In the

Regulation of Nanomaterials. Journal of Nanoparticle Research, 2011 doi: 10.1007|s11051‐011.5234.1

2. SW Bennett and AA Keller. Photocatalytic behavior of metal oxide nanomaterials in natural waters. Applied Catalysis B: Environmental, 2011, 102 (3‐4) 600‐607. doi: 10.1016/j.apcat6.2010.12.045

3. Bernhardt, E.S., Colman, B.P., Cardinale, B.J., Nisbet, R.M., Richardson C., and Yin, L. Emerging environmental crisis or part of the Green Revolution: the challenge of providing ecological predictions about nanomaterial impacts on the environment. Journal of Environmental Quality, 2010, 39: 1954‐1965

4. Chowdhury, I., Hong, Y., Walker, S. L. Container to Characterization: Impacts of Metal Oxide Handling, Preparation and Solution Chemistry on Particle Stability, Colloids and Surfaces A: Physicochemical and Engineering Aspects. 2010. 91‐95. doi: 10.10161j.colsurta2010.07.019

5. Joseph Conti, Terre Satterfield, Barbara Herr Harthorn, Nick Pidgeon “Vulnerability and Social Justice as Factors in Emergent US Nanotechnology Risk Perceptions” Risk Analysis, 2011, 31(4) online. doi: 10.1111/j.1539‐6924.2011.01608x.

6. Damoiseaux R, George S, Li M, Pokhrel S, Ji Z, France B, Xia T,Suarez E, Rallo R, Maedler L, Cohen Y, Hoek EMV and Nel A., No time to lose ‐ high throughput screening to assess nanomaterial safety (Review Article). Nanoscale, 2011. DOI: 10.1039/c0nr00618a.

7. X. Fang, R. Yu, B. Li, P. Somasundaran, K. Chandran. “Stresses Exerted by ZnO, CeO2 and Anatase TiO2 Nanoparticles on N. Europaea”. Journal of Colloid and Interface Sciences, 2010, 348 (2), p. 329 – 334. doi:10.1018/j.jcis.2010.04.075

8. X. Fang, B. Li, I. Chernyshova, P. Somasundaran. “Ranking of as received micro/nano particles by their surface energy Values at Ambient Conditions”. Journal of Physical Chemistry C, 2010, 114 (36), pp 15473–15477, doi: 10.1021/jp105720z

9. Ge, Y.; Schimel, J. P.; Holden, P. A. Evidence for negative effects of TiO2 and ZnO nanoparticles on soil bacterial communities. Environmental Science and Technology. 2011, 45, 1659‐1664, DOI: 10.1021/es103040t.

10. Saji George, Tian Xia, Robert Rallo, Yan Zhao, Zhaoxia Ji, Xiang Wang, Haiyuan Zhang, Bryan France, David Schoenfeld, Robert Damoiseaux, Rong Liu, Shuo Lin, Kenneth A Bradley, Yoram Cohen, André E Nel. Use of a High‐throughput Screening Approach Coupled with In Vivo


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Zebrafish Embryo Screening to Develop Hazard Ranking for Engineered Nanomaterials. ACS Nano, 2011, 5(3), 1805‐1817, DOI: 10.1021/nn102734s.

11. Allison M. Horst, Andrea C. Neal, Randall E. Mielke, Patrick R. Sislian, Won Hyuk Suh, Lutz Madler, Galen D. Stucky, and Patricia A. Holden, Dispersion of TiO2 nanoparticle agglomerates by Pseudomonas aeruginosa, Applied and Environmental Microbiology 2010, 76 (21), 7292‐7298. DOI: 10.1128/AEM.00324‐10

12. Zhaoxia Ji, Xue Jin, Saji George, Tian Xia, Huan meng, Xiang Wang, Elizabeth Suarez, Haiyuan Zhang, Eric MV Hoek, Hilary Godwin, Andre E. Nel, and Jeffrey I. Zink. Dispersion and Stability of TiO2 Nanoparticles in Cell Culture Media. Environmental Science & Technology, 2010, 44(19), 7309‐7314. doi:10.1021/es100417s.

13. Jiang, XM; Brinker, CJ. Rigid templating of high surface‐area, mesoporous, nanocrystalline rutile using a polyether block amide copolymer template, Chemical Communication, 46, 6123‐6125, 2010. doi: 10.1039|c0cc01394c

14. Minghua Li, Suman Pokhrel, Xue Jin, Lutz Mädler, Robert Damoiseaux, Eric M.V. Hoek. Stability, Bioavailability, and Bacterial Toxicity of ZnO and Iron‐Doped ZnO Nanoparticles in Aquatic Media, Environmental Science & Technology, 2011, 45 (2) 755‐761, DOI: 10.1021/es102266g

15. Rong Liu, Robert Rallo, Saji George, Zhaoxia Ji, Sumitra Nair, Andre´ E. Nel, and Yoram Cohen. Classification nano‐SAR modeling for the cytoxicity of a family of Metal Oxide Nanoparticles. SMALL. 2011, online, DOI: 10.1002/smll.201002366. (P)

16. López‐Moreno, M.L., de la Rosa, G., Hernández‐Viezcas, J.A., Castillo‐Michel, H. Peralta‐Videa, J.R., Gardea‐Torresdey, J.L. 2010. Evidence of the differential biotransformation and genotoxicity of ZnO and CeO2 nanoparticles on soybean (Glycine max) plants. Environmental Science and Technology, 44(19), pp.7315‐7320, 2010. doi:10.1021/es903891g

17. López‐Moreno, M.L., de la Rosa, G., Hernández‐Viezcas, J.A, Peralta‐Videa, J.R., Gardea‐Torresdey, J.L. XAS corroboration of the uptake and storage of CeO2 nanoparticles and assessment of their differential toxicity in four edible plant species. Journal of Agricultural and Food Chemistry 2010, 58 (6), 3689‐3693. doi: 10.1021/jf904472e.

18. Timothy F. Malloy, Disruption Conventional Policy: The Three Faces of Nanotechnology, UCLA Journal of Environmental Law & Policy , 2010, 28, 1‐6.

19. C. Marambio‐Jones, E.M.V. Hoek. A review of the antibacterial effects of silver nanomaterials and potential implications for human health and the environment. Journal of Nanoparticle Research 2010, 12:1531‐1551, March 2010. doi: 10.1007/s1 1051.010.9900.y

20. Robert J. Miller, Hunter S. Lenihan, Erik B. Muller, Nancy Tseng, Shannon K. Hanna, and Arturo A. Keller, Impacts of metal oxide nanoparticles on marine phytoplankton. Environmental Science & Technology. 2010, 44(19), 7329 – 7334. doi: 10.1021/es100247x.


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21. Peralta‐Videa, J.R., Zhao, L., Lopez‐Moreno, M.L., de la Rosa, G., Hong, J., Gardea‐Torresdey, J.L. Nanomaterials and the environment: A review for the biennium 2008‐2010. Journal of Hazardous Materials 2011, 186, 1‐15. doi: 10.1016/j.jhazmat.2010.11.020.

22. Robert Rallo, Bryan France, Rong Liu, Sumitra Nair, Saji George, Robert Damoiseaux, Francesc Giralt, Andre Nel, Kenneth Bradley and Yoram Cohen. Self‐Organizing Map Analysis of Toxicity‐Related Cell Signaling Pathways for Metal and Metal Oxide Nanoparticles. Environmental Science and Technology. 2011, 45, 1695‐1702, DOI: 10.1021/es103606x.

23. Rico C. M., Majumdar, S., Duarte‐Gardea, M., Peralta‐Videa, J.R. Gardea‐Torresdey, J.L. Interaction of nanoparticles with edible plants and their possible implications in the food chain. Journal of Agricultural and Food Chemistry . 2011, online, doi: 10.1021/jf104517j

24. Somasundaran, P., X. Fang, S. Ponnurangam, B. Li, Nanoparticles: Characteristics, Mechanisms, and Modulation of Biotoxicity, KONA powder and particle journal, 2010, 28, 38‐49.

25. Thio, BJ, Lee, J, Meredith, C, Keller, AA. Measuring the influence of solution chemistry on the adhesion of Au nanoparticles to mica using colloid probe atomic force microscopy. Langmuir, 2010, 26 (17), pp 13995–14003

26. Thio, BJ, Zhou, DX, Keller, AA. Influence of natural organic matter on the aggregation and deposition of titanium dioxide nanoparticles onto silica. J. Hazardous Materials. 2011, online, doi: 10.1016/j.jhazmat.2011.02.072.

27. Courtney R. Thomas, Saji George, Allison M. Horst, Zhaoxia Ji, Robert J. Miller, Jose R. Peralta‐Videa, Tian Xia, Lutz Mädler, Jorge L. Gardea‐Torresdey, Patricia A. Holden, Arturo Keller, Hunter S. Lenihan, Jeffrey I. Zink, Andre E. Nel, Nanomaterials in the environment: from materials to high throughput screening to organisms, ACS Nano(Focus Review), 5(1), 13‐20, 2011. DOI: 10.1021/nn1034857

28. Xiang Wang, Tian Xia, Susana Addo Ntim, Zhaoxia Ji, Saji George, Huan Meng, Haiyuan Zhang, Vincent Castranova, Somenath Mitra, André E. Nel, Quantitative techniques for assessing and controlling the dispersion state and biological effects of multi‐walled carbon nanotubes in mammalian tissue culture cells, ACS Nano, 4(12), 7241‐7252, 2010. doi: 10.1021/nn1021126.

29. Rebecca Werlin, John H. Priester, Randall E. Mielke, Stephan Krämer, Susan Jackson, Peter K. Stoimenov, Galen D. Stucky, Gary N. Cherr, Eduardo Orias, and Patricia A. Holden. Biomagnification of cadmium selenide quantum dots in a simple experimental microbial food chain, Nature Nanotechnology 6, 65‐71 (2011). DOI: 10.1038/NNANO.2010.251

30. Worsley, K. A. ; Kalinina, I.; Bekyarova, E.,; Haddon, R. C.; Functionalization and Dissolution of Nitric Acid Treated Single‐Walled Carbon Nanotubes. Journal of American Chemical Society, 2009, 131 (50), pp 18153–18158, doi: 10.1021/ja906267g

31. Tian Xia, Yan Zhao, Tina Sager, Saji George, Suman Pokhrel, Ning Li, David Schoenfeld, Huan Meng, Sijie Lin, Xiang Wang, Meiying Wang, Zhaoxia Ji, Jeffrey I. Zink, Lutz Mädler, Vincent Castranova, Shuo Lin, Andre E. Nel, Decreased dissolution of ZnO by iron doping yields


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nanoparticles with reduced toxicity in the rodent lung and zebra fish embryos, ACS Nano, 2011, 5(2), 1223‐1235. DOI: 10.1021/nn1028482.

32. Haiyuan Zhang , Tian Xia , Huan Meng , Min Xue, Saji George, Zhaoxia Ji , Xiang Wang , Rong Liu, Meiying Wang , Robert Rallo , Robert Damoiseaux , Yoram Cohen, Kenneth A. Bradley, Jeffrey I. Zink, Andre E. Nel. Differential toxicological effects of cationic mesoporous silica nanoparticles (MSNP) in undifferentiated and differentiated bronchial epithelial cells. ACS Nano. March 2, 2011 online. DOI: 10.1021/nn200328m.

33. Zhou, D., Keller, A., Role of morphology in the aggregation kinetics of ZnO nanoparticles. Water Research, May 2010, 44, (9), 2948 ‐ 2956.

Leveraged Publications ‐ Journals 34. Arias, J., Peralta‐Videa, J.R., Ellzey, J.T., Viveros, M.N., Ren, M., Mokgalaka‐Matlala, N.S., Castillo‐

Michel, H., Gardea‐Torresdey, J. L. Plant growth and metal distribution in tissues of Prosopis juliflora‐velutina grown on chromium contaminated soil in the presence of Glomus deserticola. Environmental Science and Technology, 2010, 44(19), 7272–7279.

35. Dumas EM, Ozenne V, Mielke RE, Nadeau, JL. Mechanisms of Toxicity of CdTe Quantum Dots in Bacterial Strains. IEEE Transactions Nanobioscience. 2009 Mar;8(1):58‐64. Epub 2009 Mar 16. PMID: 19304497.

36. Gonzalez, C.M., Hernandez, J., Parsons, J.G., Gardea‐Torresdey, J. L. Removal of selenite and selenate from aqueous solutions using a magnetic iron/manganese oxide nanomaterial. Microchemical Journal 2010, 96(2), 324‐329.

37. Gonzalez, H.O., Hu, J., Gaworecki, K.M., Roling, J.A., Baldwin, W.S., Gardea‐Torresdey, J.L., Bain, L.J. Dose‐Responsive Gene Expression Changes in Juvenile and Adult Mummichogs (Fundulus heteroclitus) After Arsenic Exposure. Marine Environmental Research, 2010, 70(2), 133‐141. doi:10.1016/j.marenvres.2010.04.003.

38. BC Heng, GK Das, X Zhao, LL Ma, T Tan, K Ng, J. Loo. Comparative cytotoxicity evaluation of lanthanide nanomaterials on mouse and human cell lines with metabolic and DNA quantification assays. Biointerphases 2010. Vol. 5 (3): FA88‐FA97.

39. Heng, B. C., X. Zhao, S. Xiong, K. Ng, F. Boey, J. Loo. "Toxicity of zinc oxide (ZnO) nanoparticles on human bronchial epithelial cells (BEAS‐2B) is accentuated by oxidative stress." Food and Chemical Toxicology 2010; 48(6): 1762‐1766. doi: 10.1016|j.fct.2010.04.023

40. Jiang XM, Y. Jiang, N. Liu, H. Xu, S. Rathod, P. Shah, C. Jeffrey Brinker, Controlled Release from Core‐Shell Nanoporous Silica Particles, Journal of Nanomaterials, 2011, online, doi: 10.1155|2011|760237

41. Lopez‐Gonzalez, H., Peralta‐Videa, J.R., Romero‐Guzman, E.T., Rojas‐Hernandez, A., Gardea‐Torresdey, J.L. Determination of the Hydrolysis Constants and Solubility Product of Chromium(III) from Reduction of Dichromate Solutions by ICP‐OES and UV–Visible Spectroscopy, Journal of Solution Chemistry, April 2010, 39 (4), 522‐532. doi: 10.1007/s10953‐010‐9522‐0.


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42. Timothy F. Malloy, Nanotechnology Regulation: A Study in Claims Making, ACS Nano 2011, 5(1), 5‐12: doi: 10.1021/nn103480e.

43. H. Meng, M. Liong, T. Xia, Z. Li, Z. Ji, J.I. Zink and A.E. Nel “Engineered Design of Mesoporous Silica Nanoparticles to Deliver Doxorubicin and P‐Glycoprotein siRNA to Overcome Drug Resistance in a Cancer Cell Line,” ACS Nano, 2010, 4, 4539‐4550.

44. Montes, M. Mayoral, A., Deepak, F.L., Parsons, J.G., Peralta‐Videa, J.R., Jose‐Yacaman, M., Gardea‐Torresdey, J.L. Anisotropic gold nanoparticles and gold plates biosynthesis using alfalfa extract. Journal of Nanoparticle Research. Web 2011, doi: 10.1007/s11051‐011‐0230.5.

45. Parsons, J.G., Lopez, M.L, Gonzalez, C., Peralta‐Videa, J.R., Gardea‐Torresdey, J.L. Toxicity and biotransformation of uncoated and coated nickel hydroxide nanoparticles on mesquite plants. Environmental Toxicology and Chemistry. 2010, 29(5), 1146‐1154. doi: 10.1002/etc.146 ISSN 0730‐7268.

46. Xiong SJ, XX Zhao, BC Heng, K Ng, J Loo. “Cellular uptake of poly‐(D, L‐lactide‐co‐glycolide) (PLGA) nanoparticles synthesized through solvent emulsion evaporation and nanoprecipitation method”. Biotechnology Journal. 2011, 6, online, DOI: 10.1002/biot.201000351.

47. Yichi Zhang, Yifeng Shi, Ya‐Hsuan Liou, April M. Sawvel, Xiaohong Sun, Yue Cai, Patricia A. Holden, Galen D. Stucky. “High Performance Separation of Aerosol Sprayed Mesoporous TiO2 Sub Microspheres from Aggregates via Density Gradient Centrifugation”, Journal of Materials Chemistry 20, 4162‐4167. 2010. DOI 10.1039/b926183d.

48. Zhao, Y., Peralta‐Videa, J. R., Lopez‐Moreno, M. L., Ren, M., Gardea‐Torresdey, J. L. Kinetin increases chromium absorption, modulates its distribution, and changes the activity of catalase and ascorbate peroxidase in Mexican Palo Verde. Environmental Science and Technology. 2011, 45, 1082‐1087. doi: 10.1021/es102647w.

Book Chapters 49. Mamadou Diallo and Jeffrey Brinker. With contributions from: André Nel, Mark Shannon, Nora

Savage, Norman Scott, James Murday. Chapter 5. Nanotechnology for Sustainability, Environment, Water, Food, and Climate. Nanotechnology Research Directions for Societal Needs in 2020. Editors: Roco M, Mirkin C, and Hersam M. Boston and Berlin. Springer.

50. Barbara Herr Harthorn. “Methodological Challenges Posed by Emergent Nanotechnologies and Cultural Values.” In The Handbook of Emergent Technologies and Social Research, Ed. Sharlene Nagy Hesse‐Biber, Oxford University Press. 2011. Chapter 3. ISBN13 : 9780195373592

51. B. Herr Harthorn. “Gender and Nanotechnology”, Encyclopedia of Nanoscience and Society, ed. David Guston. Sage Publications, July 2010. ISBN 9781412969871

52. B. Herr Harthorn. “Risk Amplification.” Encyclopedia of Nanoscience and Society, ed. David Guston. Sage Publications, July 2010. ISBN 9781412969871

53. B. Herr Harthorn. “Risk Attenuation.” Encyclopedia of Nanoscience and Society, ed. David Guston. Sage Publications, July 2010. ISBN 9781412969871


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54. Chad A. Mirkin, Andre Nel, C. Shad Thaxton. With contributions from: Barbara A. Baird, Carl Batt,

David Grainger, Sanjiv Sam Gambhir, Demir Akin, Otto Zhou, J. Fraser Stoddart, Thomas J. Meade, Piotr Grodzinski, Dorothy Farrell, Harry F. Tibbals, Joseph De Simone. Chapter 7. Applications: Nanobiosystems, Medicine, and Health. Nanotechnology Research Directions for Societal Needs in 2020. Editors: Roco M, Mirkin C, and Hersam M. Boston and Berlin. Springer.

55. André Nel, David Grainger, Pedro Alverez, Santokh Badesha, Vincent Castranova, Mauro Ferrari, Hilary Godwin, Piotr Grodzinski, Jeff Morris, Nora Savage, Norman Scott, Mark Wiesner. Chapter 4. Nanotechnology Environmental, Health and Safety Issues. Nanotechnology Research Directions for Societal Needs in 2020. Editors: Roco M, Mirkin C, and Hersam M. Boston and Berlin. Springer.

56. Mihail Roco, Barbara Herr Harthorn, David Guston & Phillip Shapira. 2011. Innovative and responsible governance of nanotechnology for societal development. Ch. 13 in Nanotechnology Research Directions for Societal Needs in 2020, ed. M. Roco. Boston and Berlin:Springer.

Conference Proceedings, Reports, and Other Articles 57. Beaudrie, C.; Emerging Nanotechnologies and Life Cycle Regulation: An Investigation of Federal

Regulatory Oversight from Nanomaterial Production to End of Life. Chemical Heritage Foundation, 53pp. 2010.

58. Saji George. Pulmonary responses to multi‐walled carbon nanotube exposure. Spheres, Winter 2010; 8(2), p.7.

59. Barbara Herr Harthorn. Public participation in nanotechnology – should we care? Op ed on 2020 Science, May 4, 2010.

60. H. Liu. Modeling aggregation and size distribution of nanoparticles with Monte Carlo simulation, Proceedings of NSTI‐NanoTech 2010, Vol. 3, 532‐534, ISBN 978‐1‐4398‐3415‐2.

61. S. Surawanvijit, M. Kim, Y. Cohen, Analysis of Membrane Filtration Efficiency in Removal of Metal Oxide Nanoparticles from Aqueous Nanoparticle Suspension in the Presence of Coagulation Pretreatment, Proceedings of NSTI‐NanoTech 2010, Vol. 3, 591‐593, ISBN 978‐1‐4398‐3415‐2

62. Tian Xia, Huan Meng, Saji George, Haiyuan Zhang, Xiang Wang, Zhaoxia Ji, Jeffrey I Zink, Andre E. Nel. Strategy for toxicity screening of nanomaterails. Materials Matters (Sigma) 2010, 5, 82‐83.

Patents There are no patentable activities to report to date 16. Biographical Information Short biographical information for new Center faculty member follows:

Mark Hersam, Northwestern University, Professor, Materials Science and Engineering

MARK C. HERSAM Materials Science and Engineering Tel: (847) 491-2696 Northwestern University Fax: (847) 491-7820 2220 Campus Drive E-mail: [email protected] Evanston, IL 60208-3108 WWW: http://www.hersam-group.northwestern.edu/ Professional Preparation University of Illinois at Urbana-Champaign Electrical Engineering B.S. 1996 University of Cambridge, UK Physics M.Phil. 1997 University of Illinois at Urbana-Champaign Electrical Engineering Ph.D. 2000

Appointments 2007-present Professor, Dept. of Chemistry, Northwestern University 2006-present Professor, Materials Science and Engineering, Northwestern University 2000-2006 Assistant Professor, Materials Science and Engineering, Northwestern Univ. Summer 1999 Research Intern, Nanoscale Science Dept., IBM T. J. Watson Research Center Summer 1996 Research Aide, Materials Science Division, Argonne National Laboratory

Publications (out of ~100 total); *indicates cover article Related to the present proposal:1. A. A. Green and M. C. Hersam, “Solution phase production of graphene with controlled

thickness via density differentiation,” Nano Letters, DOI: 10.1021/nl902200b, published online ASAP on September 25, 2009.

2. *A. A. Green and M. C. Hersam, “Processing and properties of highly enriched double-wall carbon nanotubes,” Nature Nanotechnology, 4, 64 (2009).

3. *A. A. Green, M. C. Duch, and M. C. Hersam, “Isolation of single-walled carbon nanotube enantiomers by density differentiation,” Nano Research, 2, 69 (2009).

4. M. C. Hersam, “Progress towards monodisperse single-walled carbon nanotubes,” Nature Nanotechnology, 3, 387 (2008).

5. A. A. Green and M. C. Hersam, “Colored semitransparent conductive coatings consisting of monodisperse metallic single-walled carbon nanotubes,” Nano Letters, 8, 1417 (2008).

Others: 6. *Q. H. Wang and M. C. Hersam, “Room-temperature molecular-resolution characterization

of self-assembled organic monolayers on epitaxial graphene,” Nature Chemistry, 1, 206 (2009).

7. *M. S. Arnold, J. Suntivich, S. I. Stupp, and M. C. Hersam, “Hydrodynamic characterization of surfactant encapsulated carbon nanotubes using an analytical ultracentrifuge,” ACS Nano, 2, 2291 (2008).

8. A. A. Green and M. C. Hersam, “Ultracentrifugation of single-walled carbon nanotubes,” Materials Today, 10, 59 (2007).

9. M. S. Arnold, A. A. Green, J. F. Hulvat, S. I. Stupp, and M. C. Hersam, “Sorting carbon nanotubes by electronic structure via density differentiation,” Nature Nanotechnology, 1, 60 (2006).

10. M. S. Arnold, S. I. Stupp, and M. C. Hersam, “Enrichment of single-walled carbon nanotubes by diameter in density gradients,” Nano Letters, 5, 713 (2005).

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Selected Honors and Awards AVS Peter Mark Award (2006); TMS Robert Lansing Hardy Award (2006); Presidential Early Career Award for Scientists and Engineers (2005); ONR Young Investigator Award (2005); ARO Young Investigator Award (2005); Alfred P. Sloan Research Fellowship (2005); NSF CAREER Award (2002); Arnold and Mabel Beckman Young Investigator Award (2001)

Synergistic Activities (1) Teacher of the Year, Department of Materials Science and Engineering, 2003, 2007, 2009 (2) Development of a new undergraduate course entitled “Nanomaterials,” 2001- (3) National Science Foundation SBIR, IMR/MRI, MRSEC, and NUE Panelist, 2001- (4) Director of the Nanoscale Science and Engineering Center REU Program, 2002- (5) Editor-in-Chief of Nanoscape (the journal of undergraduate research in nanoscience), 2003-

Collaborators & Other Affiliations (i) Collaborators Michael Bedzyk (Northwestern); Jeffrey Elam (Argonne); Graham Fleming (Berkeley); Franz Geiger (Northwestern); Henri Happy (CNRS); Achim Hartschuh (Munich); Ladislav Kavan (J. Heyrovsky Institute); Tobin Marks (Northwestern); SonBinh Nguyen (Northwestern); Michael Pellin (Argonne); Tamar Seideman (Northwestern); Peter Stair (Northwestern); Samuel Stupp (Northwestern); Richard Van Duyne (Northwestern); Bruce Weisman (Rice)

(ii) Graduate and Professional Advisors M.Phil. Advisor - Prof. Mark Welland, Engineering Department, Cambridge University Ph.D. Advisor - Prof. Joseph Lyding, Elec. Eng. Dept., Univ. of Illinois at Urbana-Champaign Intern Advisor - Dr. Phaedon Avouris, Nanoscale Science Dept., IBM Watson Research Center

(iii) Thesis Advisor and Postgraduate-Scholar Sponsor Postgraduate-Scholars (11): Lixin Cao (2002-03) Chinese Acad. Sci. Alexander Manasson (2007-08) Eriktron Edward Foley (2001-present) Adam McFarland (2004-05) Eli Lilly Nathan Guisinger (2005-06) Argonne Gordana Ostojic (2005-present) Md. Zakir Hossain (2009-present) Vinod Sangwan (2009-present) John Ireland (2004-2008) NanoInk John Tovar (2003-05) Johns Hopkins Don Kramer (2002-05) Smith & Nephew

Graduate Students (26): Justice Alaboson (2007-present) Joshua Keller (2004-present) Michael Arnold (PhD, 12/06) U. of Wisc. Reagan Kinser (PhD, 12/05) TRW Andrew Baluch (MS, 12/04) Foley & Lardner Joe Lee (MS, 6/08) IBM Rajiv Basu (PhD, 12/06) Intel Ben Leever (2005-present) Steven Christensen (PhD, 12/08) Argonne Tony Liang (2008-present) David Comstock (2003-present) Albert Lipson (2007-present) Norma Cortes (2004-present) Liam Pingree (PhD, 12/06) Boeing Matthew Duch (2007-present) Matthew Schmitz (MS, 12/07) USMC Ken Everaerts (2008-present) Matthew Such (MS, 12/02) USPTO Alex Green (2005-present) Timothy Tyler (2006-present) Mark Greene (PhD, 12/05) NIST Michael Walsh (2004-present) Nathan Guisinger (PhD, 12/05) Argonne Qing Hua Wang (2005-present) Hunter Karmel (2006-present) Nathan Yoder (PhD, 12/07) NanoIntegris

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Table 6: Partnering Institutions - Draft ReportNSEC Center: CEIN: Predictive Toxicology Assessment and Safe Implementation of Nanotechnology in the Environment

Institution Type Name of Institution

ReceivesFinancialSupport

From Center

ContributesFinancial

Support ToCenter

MinorityServing

InstitutionPartner

FemaleServing

InstitutionPartner

NationalLab/ Other

Govt.Partner

IndustryPartner

MuseumPartner

InternationalPartner

I. AcademicPartneringInstitution(s)

Cardiff University Y

Centro de Investigacion yde Estudios Avanzados

del Instituto PolitechnicnoNactional (CINVESTAV)

Y

Columbia University Y

Instituto Nacional deSalud Publica Y

Nanyang TechnologicalUniversity Y

Universitat Rovira I Virgili Y

University College Dublin Y

University of Bremen Y Y

University of BritishColumbia Y Y

University of California,Davis Y

University of California,Riverside Y Y

University of California,Santa Barbara Y Y

University of New Mexico Y Y

University of Texas, ElPaso Y Y

Total Number ofAcademicPartners

14 8 1 3 0 0 0 0 8

II. Non-academicPartneringInstitution(s)

California Science Center Y

Lawrence BerkeleyNational Laboratory Y

Lawrence LivermoreNational Laboratory Y

Sandia NationalLaboratory Y

Santa Monica PublicLibrary Y

Total Number ofNon-academicPartners

5 0 0 0 0 3 0 2 0

1 of 1 4/12/2011 3:46 PM

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