NC_old1194: Nanotechnology and Biosensors

(Multistate Research Project)

Status: Inactive/Terminating

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SAES-422 Reports

Annual/Termination Reports:

[02/13/2018] [11/05/2018] [06/06/2019] [07/24/2020] [03/27/2022]

Date of Annual Report: 02/13/2018

Report Information

Annual Meeting Dates: 05/17/2017 - 05/19/2017
Period the Report Covers: 10/01/2016 - 09/30/2017

Participants

Sundaram Gunasekaran (University of Wisconsin), Zhongyang Cheng (Auburn University), Yanbin Li (University of Arkansas), Jeremy Tzeng (Clemson University), Anye Wamucho (University of Kentucky), Paul Takhistov (Rutgers), Margaret Frey (Cornell University), Vince Bralts (advisor, Purdue University), Mengshi Lin (University og Missouri, Columbia), Evangelyn Alocilja (Michigan State University), Carmen Gomes (Texas A&M University), Eric McLamore (University of Florida), Renita Horton (Mississippi State University), Mingming Wu (Cornell University), Ramaraja Ramasamy (University of Georgia), Chenxu Yu (Iowa State University) and Jonathan Claussen (Iowa State University).

Brief Summary of Minutes

The minutes of the 2016 annual meeting were approved. Vince Bralts provided administrative update. NC 1194 was renewed in 2016, until 2021. Vince is in partial retirement. He will be identifying a candidate to replace him when he retires. To prepare for the renewal in year 2020, we need to gather information along the way. Station reports are critical.

The officers for 2017-18 are as follows: Jenna Rickus, Chair (moves up from vice-chair); Jeremy Tzeng, Vice-Chair (moves up from Secretary), and Mengshi Lin was elected as Secretary.

The group decided to hold the 2018 meeting in conjunction with Gordon Conference on Nano-Enabled Technologies to Improve Efficiency, Quality, and Health in Food and Agriculture held during June 3-8 at Mount Holyoke College, South Hadley, MA. The participants are encouraged to actively seek opportunities to present their work at this conference.

The following are the brief highlights of the station reports:

Alocilja (MSU): Is a part of a Global Alliance. The idea is to share the learning experience, i.e., peer-to-peer learning system. Have funding from the Philippines.

Takhistov (NJ, Rutgers): Reported on a sensing technology in equine industrial (horses) and packaging materials integrated antimicrobial with sensors for automatic release.

Li (AR): Received grants from Walmart to study supply chain. Three professors from three universities in China are participating in this study. The focus is on how to get biosensors breeder, farm, risk assessment (cannot shut down the farms based on one contaminant).

Lin (MO): Developing sensors for detecting pesticide residues in fruits, fruit juice, milk etc. using gold nanorods and nanocomposites. The detection limit is 500-600 ppb. Cellulose nanosubstrate is used for the detection of pesticides on apple.

Wamucho (KY): Studying the toxicity of nanoparticles (silver, titanium) using elegans.

Ramasamy (GA): Developing Enzyme-nanocomposite based biosensors to detect stress-released chemical compounds in plants. Also interested in foodborne pathogens using impedance based biosensor.

Frey (Cornel): Synthesizing surface functionalized nanofiber. Interested in detecting and removing pollutants in wastewater treatment facility.

Tzeng (Clemson): Reported the work on detecting and monitoring implant infection with X-ray Excited luminescence chemical imaging.

Gunasekaran (WI): Synthesizing various nanomaterials and developing biosensors for detecting microorganisms and heavy metals present in food and water.

Horton (MS): Using a microfluidic device to study cardiovascular diseases. Also interested in using nanofiber.

Yu and Claussen (IA) presented their work at the NIFA grantees session.

Bralts, Alocilja, Bhalearo, and Claussen presented their research in the USDA Special Sessions on Nanobiomaterials in Food & Agriculture at the Nanotech 2017 meeting during May 14-17, just prior to the NC1194 meeting.

Discussions, Suggestions, Recommendations:

Alocilja: How to pool the expertise of NC1194 together to better articulate a vision and mission for us to define the field. We should set some well-defined goals to improve our visible impacts. Perhaps focus on Listeria detection or water contamination, to focus our efforts so that NC 1194 members can work collaboratively.

The following major problems (members who are interested in) were identified:

Inline measurement processing (Yu*, Lin, Li, Ramasamy, and Takhistov)

Matrix challenge (Alocilja*, Gunasekaran, Ramasamy, Frey, and Takhistov)

False-positive and false-negatives (Ramasamy*, Alocilja, and Gunasekaran,)

Live vs. dead cells detection (Tzeng*, Ramasamy*, Alocilja, and Wamucho (Tsyusko lab))

The lead member in each, identified with *, will follow-up with discussions and report at the 2018 meeting.

Accomplishments

This is a report of accomplishment of NC1194 for the period October 1, 2016 through September 30, 2017 from participating institutions from different states. The findings have been disseminated to the scientific community via seminars, national/international conferences, manuscripts, and websites. The objectives of this project are: <ol> <li>Develop new technologies for characterizing fundamental nanoscale processes</li> <li>Construct and characterize self-assembled nanostructures</li> <li>Develop devices and systems incorporating microfabrication and nanotechnology</li> <li>Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems</li> <li>Develop/improve education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment</li> <li>Improve academic-industry partnership to help move the developed technologies to commercialization phase</li> </ol> Station: Michigan State University PI: Evangelyn Alocilja Objective(s) Addressed: #3 Summary of Work: <ul> <li>Our nano-biosensing technologies continue to be validated in clinical and biological samples (human, animal, and plant) for rapid disease and microbial-contaminant detection in our lab at MSU as well as with our collaborators around the</li> <li>We are developing new antimicrobial nanoparticles and antimicrobial films to help reduce foodborne</li> <li>Our technology on nanoparticle-based anti-counterfeiting devices is continually featured in the Science of Innovation educational program by the National Science Foundation, US Patent and Trademark Office, and NBC Learn as a national resource to encourage and recruit K-12 students to the science fields. The video is entitled "Science of Innovation: Anti- Counterfeiting Devices" and can be viewed at <a href="http://www.nbclearn.com/innovation/cuecard/62970">nbclearn.com/innovation/cuecard/62970.</a> This material will impact thousands of K-12 students and teachers not only in the US but also around the world.</li> <li>My TEDMED talk on nano-biosensors continues to gain audiences from many sectors. The TED talk is featured in the following website: <a href="http://www.youtube.com/watch?v=QGauiO0Eev0">http://www.youtube.com/watch?v=QGauiO0Eev0</a>.</li> <li>Our publications and conference presentations allowed the dissemination of our research work to a broader group of researchers and potential users both in the US and around the world.</li> <li>Technology transfer was our continuing activity. We received two US patents and filed several new invention disclosures while improving the technologies that are being reviewed by the US Patent and Trademark</li> <li>We have signed Confidential Disclosure Agreements (CDA) with two private companies to explore commercialization of our</li> <li>We have trained 13 undergraduate students, 2 PhD students, 1 high school student, and 1 high school teacher on nanotechnology and biosensors. The students won three awards. We have also trained two scientists from the Philippines, two scientists from Peru, two scientists from Nepal, and one medical professional from India on the use of our technologies. These students and scientists will become the future research leaders in the emerging field of nano-biosensing for global health, biodefense, food safety, water quality, and product</li> </ul> Station: University of Arizona PI: Jeong-Yeol Yoon Objective(s) Addressed: #3 Summary of Work: There is a growing need to develop a handheld, smartphone-based biosensor that can detect the type and concentration of pathogens from myriads of food (fresh produce and meat) and water (waste and irrigation) samples, as well as urine, blood, and tissue samples from animal and human subjects. These biosensors must be designed and manufactured to be easy-to-use, all-in-one, and extremely sensitive (down to single cell level or picogram protein level). What has been done? <ol> <li>Smartphone detection system has further been improved, with sophisticated image processing algorithms and smartphone-based fluorescent microscope, for detecting pathogens and environmental toxicants.</li> <li>Smartphone- and paper-based organ-on-a-chip system has newly been developed and used as a mimic for human kidney and liver, for quantifying the effects of environmental toxicants.</li> <li>An angular photodiode array system was designed and fabricated to instantly analyze the bacterial infection on porcine skin, without using any chemicals or reagents.</li> </ol> Impacts: <ul> <li>Our smartphone-based handheld biosensors can further be tested to detect virtually any types of water samples at much lower cost (<$10 per assay).</li> <li>Our smartphone-based organ-on-a-chip (OOC) device can be used for screening myriads of water samples. The same device can also be utilized for evaluating the toxicity of commercial drugs, which will significantly lower the time and labor necessary for a series of laboratory tests, animal tests, and human trials.</li> <li>Both devices can significantly save the cost, time, and effort necessary to conduct conventional assays. In addition, both devices can be used in field, greatly reducing the sample-to-answer time from a couple of days to less than 10 min, protecting the general public from potential health risks from water and environment.</li> </ul> Station: FL (University of Florida) PI: Bin Gao Objective(s) addressed: #1, #2 Summary of Work: In addition to develop new technologies to characterize and understand fundamental nanoscale processes, we have also explored the environmental applications and implications of nanotechnology. The project has provided training and professional development opportunities to graduate and undergraduate students. The results have been published in several peer-reviewed journal articles and been presented in professional meetings and conferences. Furthermore, the findings from the research project was also integrated with education by training graduate and undergraduate students with a diverse array of backgrounds. Outputs: <ul> <li>H. Lyu, B. Gao, F. He, C. Ding, J.C. Tang, J.C. Crittenden. 2017. Ball-milled carbon nanomaterials for energy and environmental applications. ACS Sustainable Chemistry & Engineering, 5 (11), 9568-9585.</li> <li>Wang, B. Gao, D.S. Tang, H.M. Sun, X.Q. Yin, C.R. Yu. 2017. Effects of temperature on graphene oxide deposition and transport in saturated porous media. J. of Hazardous Materials, 331, 28-35.</li> <li>Wang, B. Gao, Y. Li, A.E. Creamer, F. He. 2017. Adsorptive removal of arsenate from aqueous solutions by biochar supported zero-valent iron nanocomposite: Batch and continuous flow tests. Journal of Hazardous Materials, 322, 172-181.</li> </ul> Station: FL (University of Florida) PI: Eric S. McLamore Investigators/Participants: Bin Gao, Bruce Welt, John Schueller, Hitomi Yamaguchi, Bernard Hauser, Hal Knowles III, Ismail Ocsoy, Nicholas Cavallaro, Ishika Khondaker Objective(s) Addressed: #2, #3, #4, #5, #6 Summary of Work: Construct and characterize self-assembled nanostructures: We published two peer reviewed conference proceedings (including a platform presentation), and submitted one patent (in review) related to development of self-actuating nanobrush/aptamer hybrid materials for sensing [1-3]. Develop devices and systems incorporating microfabrication and nanotechnology. We developed value added nanotechnology products from agricultural waste for food packaging, solar cells, sensors [4-5], as well as new sensor systems for studying signaling in plant/mammalian systems [6-8]. Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems. Together with indigenous communities, we used commercial sensors for studying mercury toxicity in rural Colombia [9] and we also developed new green synthesis methods for fabricating nanoparticles to be used in sensing applications within the agricultural/food industry [10-11]. Finally, we demonstrated a chemical-free nanoscale modification of pipes used to transport milk in a dairy farm based on mechanical abrasion [12] Develop/improve education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment. We held workshops and disseminated training manuals with high school teachers in the USA (Florida, Maryland) and also abroad (Colombia, China) for creating flexible graphene circuits [13-16]. Improve academic-industry partnership to help move the developed technologies to commercialization phase. Our group in Florida initiated academia-industry projects with AES Controls on Iot and cybersecurity for food safety monitoring, and also with BRIDG for establishing a work group to create a draft national nanotechnology roadmap for smart ag/food. In addition to these activities, we published an invited critical/comprehensive review of food safety sensors in the IFT journal, contributing to translation of nanoscale research into industry products [17] as well as a review article resulting from a NSF/USDA workshop on the food/energy/water nexus together with industry partners [18]. Outputs (Referenced in above text): <ul> <li>Althawab, S., Oliveira, D. A., Smith, C., Cavallaro, N., McLamore, E.S., Gomes, C. (2017) Label-free, rapid Listeria monocytogenes biosensor based on a stimulus response nanobrush and nanometal hybrid electrode. Proceedings of the Tech Connect Nanotechnology Conference. vol. 3: pp. 279-282</li> <li>McLamore, E.S., I. Khondaker, C. Gomes, D. Alves De Oliveira (2017). Stimulus Response Biosensor for Determining Bacteria Viability Using Lectin-Glycoenzyme Nanobrushes. Proceedings of the Tech Connect Nanotechnology Conference. Vol. 3: 991-999.</li> <li>Khondaker, I., E.S. McLamore (2017). Determination of Bacteria Viability By Measuring Transient Biogenic Amine Production. Patent Filed on 06/06/17; T2315-22653US00</li> <li>Demirbas, A., K. Groszman, M. Pazmino, R. Nolan, D.C. Vanegas, B. Welt, J.C. Claussen, J. Hondred, E.S. McLamore (2018) Cryoconcentration of bioflavonoid extract for enhanced photovoltaics and pH sensitive thin films. Biotechnology Progress, in press</li> <li>Demirbas, A., Y. Yagiz, Z. Boz, B.A. Welt, E.S. McLamore, W. Pelletier, S. Amarat, M. Marshall (2017) Effect of red cabbage extract on minced Nile perch fish patties vacuum packaged in high and low oxygen barrier films. Journal of Applied Packaging Research. 9(2): 35-46.</li> <li>Chaturvedi, P., D.C. Vanegas, J. Foster, B.A. Hauser, M.S. Sepulveda, E.S. McLamore (2017) Microprofiling real time nitric oxide flux for field studies using a stratified nanohybrid carbon-metal electrode. Analytical Methods. 9: 6061-6072.</li> <li>Cannon, A.E., D.C. Vanegas, J. Wang, G. Clark, E.S. McLamore, S.J. Roux. Polarized Distribution of Extracellular Nucleotides Promotes Gravity-Directed Polarization of Development in Spores of Ceratopteris richardii. Plant Journal, In review</li> <li>Yan, S., S. Dong, E.S. McLamore, T. Zhang, N. Wang, H. Yao, Y. Shen (2017) Insect Herbivory Affects the Auxin Flux Along Root Apices in Arabidopsis thaliana. J. of Plant Growth Regulation. 1-9.</li> <li>Vélez-Torres, I., D. Vanegas , E.S. McLamore, D. Hurtado (2018). Unfolding the Impacts of Mercury Usage in Artisanal Gold Mining: Women’s perspective on Environmental Conflicts in Alto Cauca, Colombia. Journal of Environment and Development, In press</li> <li>Ocsoy, I., A. Demirbas, E.S. McLamore, B. Altinsoy4, N. Ildiz, A. Baldemir (2017). Green hydrothermal synthesis of silver nanoparticles with enhanced antimicrobial activity against bacterial and fungal pathogens. Journal of Molecular Liquids, 238: 263-269.</li> <li>Ocsoy, I., S. Yusufbeyoglu, V. Yilmaz, E.S. McLamore, N. Ildız, A. Ülgen (2017). DNA Aptamer Functionalized Gold Nanostructures for Molecular Recognition and Photothermal Inactivation of Methicillin-Resistant Staphylococcus aureus. Colloids and Surfaces B: Biointerfaces. 159: 16-22.</li> <li>Ihara, I., E. Nakano, E.S. McLamore, J.K. Schueller, K. Toyoda, K. Umetsu, H. Yamaguchi (2017). Cleanability of Milk Deposits on Inner Stainless Steel Tubing Surfaces Prepared by Magnetic Abrasive Finishing. Engineering in Agriculture, Environment and Food, 10(1): 63-68</li> <li>McLamore, E.S. (2017) Nanobiosensor training workshop: creating flexible graphene circuits with a low cost laser system. Gainesville, Florida. No. of participants = 56</li> <li>McLamore, E.S. (2017) Nanobiosensor training workshop: creating flexible graphene circuits with a low cost laser system. Baltimore, Maryland. No. of participants = 18</li> <li>McLamore, E.S. (2017) Nanobiosensor training workshop: creating flexible graphene circuits with a low cost laser system. Cali, Colombia. No. of participants = 14</li> <li>McLamore, E.S. (2017) Nanobiosensor training workshop: creating flexible graphene circuits with a low cost laser system. Beijing, China. No. of participants = 22</li> <li>Vanegas, D.C. J.C. Claussen, C. Gomes, E.S. McLamore (2017) Emerging technologies for rapid monitoring of bacteria and bacterial biomarkers in food. Comprehensive Reviews in Food Science and Food Safety. 16(6): 1188–1205.</li> <li>Castell-Perez, E., C. Gomes, J. Tahtouh, R. Moreira, E.S. McLamore, H. Knowles III (2017). Food Processing and Waste within the Nexus Framework. Current Sustainable Renewable Energy Reports, 4(3): 99-108.</li> </ul> Station: Hawaii (University of Hawaii) PI: Daniel M. Jenkins Objective(s) Addressed: #6 Summary of Work: To support other research by collaborators in other states, we have also developed an open-source, potentiostat device. The device was designed to be affordable, handheld, and wireless (Bluetooth and WiFi) to facilitate adoption for diagnostic technologies in the field or on-line in processing environments, but with performance to support high quality, sensitive measurements. The hardware is capable of any standard voltammetric or amperometric technique (i.e., cyclic voltammetry and differential pulse voltammetry), and to the knowledge of the authors it is the smallest and definitely the most affordable instrument capable of conducting electrochemical impedance spectroscopy (EIS). We are currently working on a new iteration of the prototype to enable EIS between 100 Hz and 1 kHz (by incorporation of an external slower clock for the built in network analyzer, and/or use of a faster Analog to Digital Converter for to extend the frequency range at the lower end of the spectrum. We are also working on upgrades to the firmware to improve signal to noise ratio (i.e. by disabling the WiFi during analytical steps, or taking a consensus approach to measurement to reject spurious noise). The hardware is interfaced to a customized Android app available freely on Android Play (https://play.google.com/store/apps/details?id=com.diagenetix.abestat&hl=en). As a service to the profession to facilitate custom hardware development and testing by other groups, we have also developed a published Android app that can be used to easily control and collect data wirelessly from bluetooth enable hardware  https://play.google.com/store/apps/details?id=com.uhmbe.DAQCTRL&hl=en). This app allows a user to connect to a remote hardware device through a Bluetooth modem. Once connected the app notifies the remote device of the connection so it can populate and configure the interface through coded commands. Available elements for the interface include 16 generic data fields which can be plotted on an interactive graph in real time, 8 configurable buttons, 4 radio groups that can each be configured with 4 radio buttons, and 8 controls to send numerical input to the remote device. User interaction with the elements in the app result in coded information sent back to the remote device (i.e. so it can recognize button presses / selections, and receive numerical input). Numerical and textual data sent to the application can be saved to a comma delimited (.csv) file, and shared by e-mail.  Outputs: <ul> <li>Jenkins, D. M., J. Reyes-de-Corcuera. 2017. Open-source Android app for facilitating customized data acquisition, visualization, and control. Presentation 1701479 at 2017 International Meeting of American Society of Agricultural and Biological Engineers, Spokane, WA.</li> <li>Jenkins, D. M. and J. Reyes-de-Corcuera. 2017. Handheld, open-source potentiostat for high-performance electrochemical analysis in the field. Presentation 1701478 at 2017 International Meeting of American Society of Agricultural and Biological Engineers, Spokane, WA.</li> </ul>  Station: IA (Iowa State University) PI: Chenxu Yu Investigators: Yu, CH; Claussen, JO. Objective(s) Addressed: #1, #2, #3 Summary of Work: Food and biosafety is one of the key national interests. Rapid threat response relies on on-site analysis that recognizes potential hazards at the earliest possible time with high fidelity. Nanotechnology plays a key role in the development of modern sensing methodologies that support rapid response yet miniaturized sensors for quick deployment. The focus of this multistate project is to incorporate nanotechnology research and biosensor development to yield novel technological breakthroughs that will facilitate the advance of technology for in-field foodborne pathogen detection. Impacts: <ul> <li>Evaluated production and presence of carbon nanoparticles in foods, and their fluorescence and bioluminescence properties. It furthered our understanding of naturally occurring nanoscale processes in food matrix which may lead to better utilization of these nano-phenomena.</li> <li>Continue to develop nano-vaccines using self-assembled nanostructures as carriers.</li> <li>Continue to work on microfluidic Raman biosensors which integrated microfluidic device with SERS imaging to achieve single-cell level detection of pathogens in water with sub-strain level specificity. We also investigate the potential of using portable Raman imaging to diagnose Chronic Downing disease in deer.</li> </ul> Station: IL (University of Illinois at Urbana-Champaign) PI: Kaustubh D. Bhalerao, Objective(s) Addressed: #1, #6 Summary of Work: Adaptable lab scale experimental platform of complex anaerobic microbial communities developed as model, self-contained ecosystem which was perturbed with controlled addition of a) carbon sources b) nanoparticles (silver, titanium, quantum dots, gold, iron oxide) c) antibiotics to create diverse colony structures and demonstrated that it is possible to quantitatively discriminate between divergent microbiomes using both genomic and single-cell phenotyping approaches with later having advantage in terms of speed, throughput, cost with comparable accuracy. The tool was integrated into CFML (Cytometric Fingerprinting & Machine Learning) package which attracted federal funding in analyzing animal fluids of economic importance (milk, semen, manure) and collaboration with US Army Center for Environmental Health Research (USACEHR) for analyzing gut microbiomes. A large, curated database of unique cytometric fingerprints of 10 Holstein healthy & 10 sick cows was created using CFML which will be key in solving $2B sub-clinical bovine mastitis problem in the US. Motivating farmers in the use of data driven and monitoring-based approaches for the proper and judicious use of antibiotics in animal agriculture. Encouraging highly qualified students primarily from traditionally under-served ethnic minority groups and economically disadvantaged backgrounds to pursue career in science. Attracting undergraduate researchers from various educational backgrounds to get exposure and hands-on experience in the problems of economic importance though the cutting-edge research tools in Biological Engineering. Outcomes: A label-free, high throughput technique, which is sensitive to spatial and temporal changes in the structure and function of microbiomes which got incorporated into the courses taught by the PI on Biological Nanotechnology and Biological Principles. A learning module scheduled to be packaged into 4-6 hours workshop that will include hands on tools to be offered in Summer 2018 for the benefit to wider research community. The publication in Bioresource Technology, one under review in Water Research and a few in preparation in addition to two collaborative publications. Numerous invited talks, seminars and colloquia at both the national and international level. Motivated talented high school students from underrepresented minorities under RAP and REU programs. Station: WI (University of Wisconsin-Madison) PI: Sundaram Gunasekaran Objective(s) Addressed: #2, #3 Summary of Work: We synthesize various nanoscale materials, characterize, and employ them in biosensors for the detection of various analytes (e.g., microorganisms, heavy metals) in food and water samples. Sensing modality include taking advantage of surface plasmon resonance, a widely used nanoscale phenomenon and electrochemical biosensing, both via immunogenic and non-immunogenic methodologies. Outputs: <ol> <li>Gong S, Chen H, Zhou X, Gunasekaran S. 2017. Synthesis and applications of MANs/poly(MMA-co-BA) nanocomposite latex by miniemulsion polymerization. R. Soc. Open Sci. 4: 170844.</li> <li>Hahn J, Kim E, You YS, Gunasekaran S, Lim S, Choi YJ. 2017. A switchable linker-based immunoassay for ultrasensitive visible detection of Salmonella in tomatoes. J. of Food Sci. (10):2321–2328.</li> <li>Wang YC, Lu L, Gunasekaran S. 2017. Bioplymer/gold nanoparticles composite plasmonic thermal history indicator to monitor quality and safety of perishable bioproducts. Biosensors & Bioelectronics 92:109-116.</li> <li>Sadak O, Sundramoorthy AK, S Gunasekaran. 2017. Highly selective colorimetric and electrochemical sensing of iron (iii) using Nile red functionalized graphene film. Biosensors & Bioelectronics 89:430-4436</li> </ol>  Station: MO (University of Missouri, Columbia) PI: Mengshi Lin Objective(s) Addressed: #1, #2, #3, #4 Summary of Work:  In this reporting period, we used new technologies to develop nanostructures and nanosubstrates for surface-enhanced Raman spectroscopy (SERS) applications. This study aimed to use cellulose nanofibers (CNF) to develop novel CNF-based nanocomposite as a SERS substrate. CNF were cationized with ammonium ions and then interacted with citrate-stabilized gold nanoparticles (AuNPs) via electrostatic attraction to form uniform nanocomposites. The CNF-based nanostructures were loaded with AuNPs that were firmly adhered on the CNF surfaces, providing a three-dimensional plasmonic SERS platform. A Raman-active probe molecule, 4-aminothiophenol, was selected to evaluate the sensitivity and reproducibility of CNF-based SERS substrate. The intensity of SERS spectra obtained from CNF/AuNP nanocomposite was 20 times higher than that from the filter paper/AuNP substrate. The SERS intensity map demonstrates good uniformity of the CNF/AuNP substrate. CNF/AuNP nanocomposites were used in rapid detection of thiram in apple juice by SERS and a limit of detection of 52 ppb of thiram was achieved. These results demonstrate that CNF/AuNP nanocomposite can be used for rapid and sensitive detection of pesticides in food products.  We also developed CNF-based substrate for rapid detection of melamine in milk by SERS. CNF served as a highly porous platform to load with gold nanoparticles (AuNPs), which can be used as a flexible SERS substrate with nanoscale roughness to generate strong electromagnetic field in SERS measurement. The CNF/AuNP substrate was characterized by UV-vis spectroscopy and electron microscopy. Milk samples contaminated by different concentrations of melamine were measured by SERS coupled with CNF/AuNP substrate. The spectral data analysis was conducted by multivariate statistical analysis (i.e. partial least squares (PLS)). Satisfactory PLS result for quantification of melamine in milk was obtained (R = 0.94). The detection limit for melamine extracted from liquid milk by SERS is 1 ppm, which meets the World Health Organization’s requirement of melamine in liquid milk. These results demonstrate that CNF/AuNP substrate has improved homogeneity and can be used in SERS analysis to for food safety applications.  This project has provided training for two doctoral students and two Master's students. We have disseminated the results to the industry and scientific communities at professional conferences such as IFT, ACS, and IAFP.

Publications

Publications are included in the summary of Accomplishments.

Impact Statements

The participants are making important contributions to all of the stated objectives. The impacts of their work are included under the summary of Accomplishments.

Date of Annual Report: 11/05/2018

Report Information

Annual Meeting Dates: 06/04/2018 - 06/06/2018
Period the Report Covers: 10/01/2017 - 09/30/2018

Participants

Tzuen-Rong Jeremy Tzeng, Clemson University - Acting Chair
Fanbin Kong, UGA - Acting Vice Chair
Daniel M. Jenkins, University of Hawaii - Acting Secretary
Steve Lommel, NCSU - Administrative Advisor
Hongda Chen, USDA-NIFA - Bioprocess Engineering and Nanotechnology
Eric McLamore, University of Florida
Jose Reyes-de-Corcuera, UGA
Jonathan Claussen, Iowa State University
Carmen Gomes, Iowa State University
Chengxu Yu, Iowa State University
Evangelyn Alocilja, Michigan State University
Paul Takhistov, Rutgers University
Sundaram Gunasekaran, University of Wisconsin Madison
Joel Pedersen, University of Wisconsin Madison
Anhong Zhou, Utah State University

Brief Summary of Minutes

Minutes Attachment:

NC-1194 Minutes-2018.pdf

Accomplishments

This is a report of accomplishment of NC1194 for the period October 1, 2017 through September 30, 2018 from participating institutions from different states. The findings have been disseminated to the scientific community via seminars, national/international conferences, manuscripts, and web sites. The objectives of this project are: 1.     Develop new technologies for characterizing fundamental nanoscale processes 2.     Construct and characterize self-assembled nanostructures 3.     Develop devices and systems incorporating microfabrication and nanotechnology 4.     Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems 5.     Develop/improve education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment 6.     Improve academic-industry partnership to help move the developed technologies to commercialization phase   Station: University of Arkansas PI: Yanbin Li Investigators/Participants: Ronghui Wang (Research Scientist), Zach Callaway (PhD student), Wenqian Wang (PhD student), Xinge Xi (PhD student), Xiaofan Yu (PhD student), America Sotero (MS student) Objective(s) Addressed: #2 & 3 Summary of Work: Various nanoscale materials, including magnetic nanoparticles, silk nanofibers, gold nanorods, nanochannels, were synthesized or fabricated, characterized, and used in development of new biosensors for the rapid detection of pathogenic bacteria, viruses, toxic agents, and antibiotics residues in agricultural and food products. Enzymes, antibodies, aptamers and DNA probes were selected as biosensing materials and electrochemical, optical, QCM transdusing methods were employed in the design of novel biosensors. The nanomaterials-based biosensors were also integrated with microfluidic chips-based sample treatment and measurement and cell phone-based data acquisition, processing, and transmission. This project provided an opportunity for training four Ph.D. students and one M.S. student in the areas of biosensors and nanotechnology. The project also enhanced our international collaboration with Zhejiang University and China Agricultural University on a project funded by Walmart Foundation. We made 15 presentations to the industry and scientific communities at professional conferences such as ASABE, IAFP, and IBE annual meetings as well as Biosensors 2018. We also published 11 articles in different journals including Biosensors & Bioelectronics, Sensors & Actuators, Journal of Food Protection, Electroanalysis, Journal of Materials Chemistry, Journal of Electroanalytical Chemistry, Journal of Biotechnology, etc.  Outputs: <ol> <li>Cao, L.L., Q. Zhang, H. Dai, Y.C. Fu, and Li. 2018. Separation/concentration-signal-amplification in-one method based on electrochemical conversion of magnetic nanoparticles for electrochemical biosensing. Electroanalysis 30(3):517-524. DOI: 10.1002/elan.201700653</li> <li>Dai, H., Y.Q. Li, Y.C. Fu, and Li. 2018. Enzyme catalysis induced polymer growth in nanochannels: A new approach to regulate ion transport and to study enzyme kinetics in nanospace. Electroanalysis 30(2):328-335 (available online Dec. 18, 2017). DOI:10.1002/elan.201700703</li> <li>Dai, H., Y.Q. Li, Q. Zhang, Y.C. Fu, and Li. 2018. A colorimetric biosensor based on enzyme-catalysis-induced production of inorganic nanoparticles for sensitive detection of glucose in white grape wine. RSC Advances 8:33960-33967. DOI: 10.1039/c8ra06347h</li> <li>Hu, Q.Q., R.H. Wang, H. Wang, M.F. Slavik and Li. 2018. Selection of acrylamide-specific aptamers by a quartz crystal microbalance combined SELEX method and their application in rapid and specific detection of acrylamide. Sensors and Actuators: B: Chemical 273:220-227. doi.org/10.1016/j.snb.2018.06.033</li> <li>Li, Z.S., G.S. Zhou, H. Dai, M.Y. Yang, Y.C. Fu, Y.B. Ying, and Li. 2018. Biomineralization-mimetic preparation of hybrid membranes with ultra-high load of pristine metal-organic frameworks grew on silk nanofibers for hazards collection in water. Journal of Materials Chemistry A 6(8):3402-3413 (published online on December 5, 2017). DOI:10.1039/C7TA06924C</li> <li>Wang, H., L.J. Wang, Q.Q. Hu, R.H. Wang, Li and M. Kidd. 2018. Rapid and sensitive detection of Campylobacter jejuni in poultry products using a nanoparticles-based piezoelectric immunosensor integrated with magnetic immunoseparation. Journal of Food Protection 81(8):1321-1330. doi:10.4315/0362-028X.JFP-17-381</li> <li>Wang, L.J., R.H. Wang, H. Wei, and Li. 2018. Selection of aptamers against pathogenic bacteria and their diagnostics application. World Journal of Microbiology and Biotechnology 34:149. doi.org/ 10.1007/s11274-018-2528-2</li> <li>Xu, C.N., L.Y. Lan, Y. Yao, J.F. Ping, Li, and Y.B. Ying. 2017. An unmodified gold nanorods-based DNA colorimetric biosensor with enzyme-free hybridization chain reaction amplification. Sensors & Actuators: B. Chemical. 273:642-648. doi.org/:10.1016/j.snb.2018.06.035</li> <li>Yu, X.F., F. Chen, R.H. Wang, and Li. 2018. Whole-bacterium SELEX of DNA aptamers for rapid detection of E. coli O157:H7 using a QCM sensor. Journal of Biotechnology 266:39-49. (available online, Dec. 22, 2017). doi.org/10.1016/j.jbiotec.2017.12.011</li> <li>Zhang, Q., L. Zhang, H. Dai, Z.S. Li, Y.C. Fu, and Li. 2018. Biomineralization-mimetic preparation of robust metal-organic frameworks biocomposites film with high enzyme load for electrochemical biosensing. Journal of Electroanalytical Chemistry 823:40-46. doi.org/10.1016/j.jelechem.2018.04.015</li> <li>Zheng, Y., G.Z. Cai, S.Y. Wang, M. Liao, Y. Li, and J.H. Lin. 2019. A microfluidic colorimetric biosensor for rapid detection of Escherichia coli O157:H7 using gold nanoparticle aggregation and smart phone imaging. Biosensors & Bioelectronics 124-125: 143-149. doi.org/10.1016/j.bios.2018.10.006</li> </ol> Impacts: These nanomaterials-based biosensors have shown their potential to be applied to the in-field or online detection of biological and chemical agents in agricultural and food products for ensuring food quality and safety. The implementation of these biosensors may provide close to real-time data in biodetection for the development of smart agricultural and food systems based on big data, machine learning and artificial intelligence.   Station: University of Arizona PI: Dr. Jeong-Yeol Yoon Objective(s) Addressed: #3 Summary of Work: There is a growing need to develop a handheld, smartphone-based biosensor that can detect the type and concentration of pathogens from myriads of food (fresh produce and meat) and water (waste and irrigation) samples, as well as urine, blood, and tissue samples from animal and human subjects. These biosensors must be designed and manufactured to be easy-to-use, all-in-one, and extremely sensitive (down to single cell level or picogram protein level). Outputs: <ol> <li>Kattika Kaarj, Patarajarin Akarapipad and Jeong-Yeol Yoon, "Simpler, Faster, and Sensitive Zika Virus Assay Using Smartphone Detection of Loop-mediated Isothermal Amplification on Paper Microfluidic Chips," Scientific Reports, 2018, 8: 12438. [Aug. 20, 2018]</li> <li>Tiffany-Heather Ulep and Jeong-Yeol Yoon, "Challenges in Paper-Based Fluorogenic Optical Sensing with Smartphones," Nano Convergence, 2018, 5: 14. [May 4, 2018]</li> <li>Katherine E. Klug, Kelly A. Reynolds and Jeong-Yeol Yoon, "A Capillary Flow Dynamics-Based Sensing Modality for Direct Environmental Pathogen Monitoring," Chemistry - A European Journal, 2018, 24(23): 6025-6029. Hot Paper. Inside Cover. Highlighted in ChemistryViews Magazine. [Feb. 5, 2018]</li> <li>Cayla Baynes and Jeong-Yeol Yoon, "µPAD Fluorescence Scattering Immunoagglutination Assay for Cancer Biomarkers from Blood and Serum," SLAS Technology (formerly JALA - Journal of Laboratory Automation), 2018, 23(1): 30-43. [Feb. 2018]</li> <li>Soohee Cho, Tu San Park, Kelly A. Reynolds and Jeong-Yeol Yoon, "Multi-Normalization and Interpolation Protocol to Improve Norovirus Immunoagglutination Assay from Paper Microfluidics with Smartphone Detection," SLAS Technology (formerly JALA - Journal of Laboratory Automation), 2017, 22(6): 609-615. [Dec. 2017]</li> <li>Robin E. Sweeney and Jeong-Yeol Yoon, "Angular Photodiode Array-Based Device to Detect Bacterial Pathogens in a Wound Model," IEEE Sensors Journal, 2017, 17(21): 6911-6917. [Nov. 1st, 2017]</li> </ol>    Station: Clemson University PI: Dr. Tzuen-Rong Jeremy Tzeng Objective(s) Addressed: #3 Summary of Work: Pathogen attachment is a complex phenomenon and vital process for successful initiation of infection in the host. Bacterial pathogens utilize two primary mechanisms to adhere onto host cells, namely carbohydrate-protein recognition and protein-protein interaction. The adhesion structures have a high degree of preference for a particular host-cell receptor. We have developed nanoparticles functionalized with specific receptors and evaluated their ability for selective binding and killing of pathogens. Outputs: <ol> <li>Revisit of wall-induced lateral migration in particle electrophoresis through a straight rectangular microchannel: Effects of particle zeta potential. Liu, Zhijian & Li, Di & Saffarian, Maryam & Tzeng, Tzuen‐Rong & Song, Yongxin & Pan, Xinxiang & Xuan, Xiangchun. Electrophoresis, (2018), 10.1002/elps.201800198.</li> <li>Multianchored Glycoconjugate‐Functionalized Magnetic Nanoparticles: A Tool for Selective Killing of Targeted Bacteria via Alternating Magnetic Fields. Raval* YS, Fellows BD, Murbach J, Cordeau Y, Mefford OT, Tzeng TJ. Advanced Functional Materials, 2017, 27 (26): 1701473</li> </ol>   Station: University of Florida PI: Bin Gao Objective(s) Addressed: #1 & 2 Summary of Work: In addition to develop new technologies to characterize and understand fundamental nanoscale processes, we have also explored the environmental applications and implications of nanotechnology. The project has provided training and professional development opportunities to graduate and undergraduate students. The results have been published in several peer-reviewed journal articles and been presented in professional meetings and conferences. Furthermore, the findings from the research project was also integrated with education by training graduate and undergraduate students with a diverse array of backgrounds.  Outputs: <ol> <li>Suthar, B. Gao. 2017. Use of nanotechnology against heavy metals present in water. In: A.M. Grumezescu, ed. Water Purification, 75-118.London, UK, Elsevier.</li> <li>Wang, B. Gao, A.R. Zimmerman, X.Q. Lee. Impregnation of multiwall carbon nanotubes in alginate beads dramatically enhances their adsorptive ability to aqueous methylene blue. Chemical Engineering Research & Design, 2018. 133, 235-242.</li> <li>Wang, B. Gao, Y. Wan. Comparative study of calcium alginate, ball-milled biochar, and their composites on methylene blue adsorption from aqueous solution. Environmental Science and Pollution Research, 2018. doi: 10.1007/s11356-018-1497-1.</li> <li>X. Sun, S.N. Dong, Y.Y. Sun, B. Gao, W.C. Du, H.X. Xu, J.C. Wu. Graphene oxide-facilitated transport of levofloxacin and ciprofloxacin in saturated and unsaturated porous media. Journal of Hazardous Materials, 2018. 348, 92-99.</li> <li>L. Gao, Y.M. Ma, Y.M. Zhou, H.H. Song, L. Li, S.H. Liu, X.Q. Liu, B. Gao, C.Z. Liu, K.P. Zhang. High photoluminescent nitrogen-doped carbon dots with unique double wavelength fluorescence emission for cell imaging. Materials Letters, 2018. 216, 84-87.</li> <li>S. Wang, Y.X. Zhou, S.W. Han, N. Wang, W.Q. Yin, X.Q. Yin, B. Gao, X.Z. Wang, J. Wang. Carboxymethyl cellulose stabilized ZnO/biochar nanocomposites: Enhanced adsorption and inhibited photocatalytic degradation of methylene blue. Chemosphere, 2018. 197, 20-25.</li> <li>Wang, B. Gao, D.S. Tang, C.R. Yu. Concurrent aggregation and transport of graphene oxide in saturated porous media: Roles of temperature, cation type, and electrolyte concentration. Environmental Pollution, 2018. 235, 350-357</li> <li>Wang, B. Gao, D.S. Tang, H.M. Sun, X.Q. Yin, C.R. Yu. Effects of temperature on aggregation kinetics of graphene oxide in aqueous solutions. Colloids and Surfaces A-physicochemical and Engineering Aspects, 2018. 538, 63-72.</li> </ol>    Station: University of Florida PI: Eric S. McLamore Investigators/Participants: Eric S. McLamore, Associate Professor, Agricultural and Biological Engineering Department, Institute of Food and Agricultural Sciences Bin Gao, Professor, Agricultural and Biological Engineering Department, Institute of Food and Agricultural Sciences Bruce Welt, Professor, Agricultural and Biological Engineering Department, Institute of Food and Agricultural Sciences John K. Schueller, Professor, Mechanical Engineering Department Objective(s) Addressed: #3, 4, 5, & 6 Summary of Work: Our research at UF in 2018 addressed objectives 3-6, as outlined below. Our station attended the 2018NC1194 meeting, as well as the Gordon Research Conference on Nanotechnology funded by NIFA. Our station also attended annual SPIE, IBE, ASABE, and IFT meetings. Research products were disseminated through training workshops in the US and abroad, conference papers/presentations and peer reviewed journal articles at all meetings. Objective 3) Develop devices and systems incorporating microfabrication and nanotechnology. We published six articles in 2018 on the development and application of smart nano/micro sensor systems. These included nanomaterials applied to commercial screen printed electrodes, sensors developed on novel flexible carbon circuits, and post hoc analysis systems using a smart phone. Objective 4) Develop a framework for economic, environmental and health risk assessment for nanotechnologies applied to food, agriculture and biological systems. Together with indigenous communities, we applied new nanosensors for studying mercury toxicity in rural Colombia and we also worked with plant physiology labs and food packaging labs to apply our sensors, establishing new stakeholders for the technologies developed by NC1194. Objective 5) Develop/improve education and outreach materials on nanofabrication, sensing, systems integration and application risk assessment. We held workshops and disseminated training manuals with high school teachers in the USA (Florida) and also abroad (Colombia) for applying sensors to measure water quality and plant health. Objective 6) Improve academic-industry partnership to help move the developed technologies to commercialization phase. Our group in Florida signed three CDA with major companies to develop and test sensor technologies as well as explore commercialization of our technologies. The McLamore lab trained 14 undergraduate students, 3 PhD students, 3 high school student, and 2 high school teachers on nanotechnology and biosensors through grants provided by USDA/NSF. Outputs: <ol> <li>Ding, S., C. Mosher, X.Y. Lee, S. Das, A. Cargill, X. Tang, B. Chen, E.S. McLamore, C. Gomes, J.M. Hostetter (2017). Rapid and Label-free Detection of Interferon Gamma via an Electrochemical Aptasensor Comprised of a Ternary Surface Monolayer on a Gold Interdigitated Electrode Array. ACS Sensors, 2(2): 210-217.</li> <li>Garland, N.T., E.S. McLamore, N.D. Cavallaro, D. Mendivelso-Perez, E.A. Smith, D. Jing, J.C. Claussen (2018). Flexible Laser-Induced Graphene for Nitrogen Sensing in Soil. Advanced Functional Materials. 10 (45): 39124–39133. DOI: 10.1021/acsami.8b10991</li> <li>Abdelbasir, S.M. S.M. El-Sheikh, V.L. Morgan, H. Schmidt, L.M. Casso-Hartmann, D.C. Vanegas, I. Velez-Torres, E.S. McLamore (2018). Graphene-anchored cuprous oxide nanoparticles from waste electric cables for electrochemical sensing. ACS Sustainable Chemical Engineering, 6(9), pp 12176–12186. DOI: 10.1021/acssuschemeng.8b02510.</li> <li>Vanegas, D.C., L. Patiño, C. Mendez, D. Alves de Oliveira, A.M. Torres, E.S. McLamore, C. Gomes (2018). Low-Cost Electrochemical Biosensor for Detection of Biogenic Amines in Food Samples. Biosensors Journal, 8(2). DOI: 10.3390/bios8020042.</li> <li>Hills, K.D., D. Alves De Oliveira, N. Cavallaro, C. Gomes, E.S. McLamore (2018). Actuation of chitosan-aptamer nanobrush borders as a mechanism for capturing pathogens. Analyst, 143: 1650-1661. DOI: 10.1039/c7an02039b.</li> <li>Rong, Y., A.V. Pardon, K.J. Hagerty, N. Nelson, S. Chi, N.O. Keyhani, J. Katz, Shoumen Datta, C. Gomes, E.S. McLamore (2018). Post hoc support vector machine learning for biosensors based on weak protein-ligand interactions. Analyst, 143, 2066-2075. DOI: 10.1039/c8an00065d.</li> <li>Vélez-Torres, I., D. Vanegas , E.S. McLamore, D. Hurtado (2018). Mercury Pollution and Artisanal Gold Mining in Alto Cauca, Colombia: Woman's Perception of Health and Environmental Impacts. Journal of Environment and Development, 27(4) 415–444. DOI: 10.1177/1070496518794796.</li> <li>Bera, T., E.S. McLamore, B. Wasik, B. Rathinasabapathi, G. Liu (2018). Identification of a maize (Zea mays L.) inbred line adapted to low‐P conditions via analyses of phosphorus utilization, root acidification, and calcium influx. J. Plant Phys., 181(2): 275-286. DOI: 10.1002/jpln.201700319</li> <li>Cannon, A.E., D.C. Vanegas, J. Wang, G. Clark, E.S. McLamore, S.J. Roux. Polarized Distribution of Extracellular Nucleotides Promotes Gravity-Directed Polarization of Development in Spores of Ceratopteris richardii. Plant Journal, In review</li> <li>Boz, Z., Welt, B.A., Brecht, J.K., Pelletier, W., E.S. McLamore, G.A. Kiker, J.E Butler (2018). Review of challenges and advances in modification of food package headspace gases. Journal of Applied Packaging Research, 10(1): 62-67.</li> </ol>   Station: Michigan State University PI: Dr. Evangelyn Alocilja, Professor, Biosystems and Agricultural Engineering Objective(s) Addressed: #3 Summary of Work: <ol> <li>Our research was Finalist for the 2017 Manufacturing Innovation Leadership Award (US national award)</li> <li>One of students received the 2017 State of Michigan Stockholm Junior Water Prize Award; student represented the State of Michigan to the US Stockholm Junior Water Prize Competition</li> <li>Technology transfer was our continuing activity. We received two US patents and filed several new invention disclosures while improving the technologies that are being reviewed by the US Patent and Trademark Office.</li> <li>Our nano-biosensing technologies continue to be validated in clinical and biological samples (human, animal, and plant) for rapid disease and microbial-contaminant detection in our lab at MSU as well as with our collaborators around the world, such as in the Philippines, Nepal, India, Peru, Colombia, Mexico, and Poland.</li> <li>Our antimicrobial films have been tested in various foods, such as lettuce, cucumber, strawberries, ground beef, salami, cheese, and chicken meat. Bacterial reduction ranges from 4 logs to 7 logs of bacterial contamination.</li> <li>Our technology on nanoparticle-based anti-counterfeiting devices is continually featured in the Science of Innovation educational program by the National Science Foundation, US Patent and Trademark Office, and NBC Learn as a national resource to encourage and recruit K-12 students to the science fields. The video is entitled "Science of Innovation: Anti-Counterfeiting Devices" and can be viewed at www.nbclearn.com/innovation/cuecard/62970. This material will impact thousands of K-12 students and teachers not only in the US but also around the world.</li> <li>My TEDMED talk on nano-biosensors continues to gain audiences from many sectors. The TED talk is featured in the following website: <a href="http://www.youtube.com/watch?v=QGauiO0Eev0">http://www.youtube.com/watch?v=QGauiO0Eev0</a>.</li> <li>Our publications and conference presentations allowed the dissemination of our research work to a broader group of researchers and potential users both in the US and around the world.</li> <li>Technology transfer was our continuing activity. We received two US patents and filed several new invention disclosures while improving the technologies that are being reviewed by the US Patent and Trademark Office.</li> <li>We have trained 16 undergraduate students, 3 PhD students, 2 high school students, and 1 high school teacher on nanotechnology and biosensors. The students won several awards. We have also trained scientists from the Philippines, Peru, Nepal, and India on the use of our technologies. These students and scientists will become the future research leaders in the emerging field of nano-biosensing for global health, biodefense, food safety, water quality, and product integrity.</li> </ol>   Station: University of Missouri, Columbia, MO PI: Mengshi Lin, Professor, Food Science Program, University of Missouri Objective(s) Addressed: # 1, 2, & 3 Summary of Work: Our objectives are to develop new technologies for characterizing fundamental nanoscale processes; construct and characterize self-assembled nanostructures; and develop devices and systems using nanotechnology. In this reporting period, we developed an environmentally friendly and cost-effective approach to synthesize green silver nanoparticles (AgNPs) from silver precursors. Green synthesis of AgNPs was accomplished using the aqueous extract of turmeric powder, in which plant biomaterials were used as a reducing as well as a capping agent. After 24 h of reaction, the yellow color of extract was changed to dark brown-reddish due to the reduction of silver ions to AgNPs. AgNPs were characterized using UV-vis spectroscopy, Fourier transform infrared spectroscopy, transmission electron microscopy (TEM), and energy dispersive X-ray spectroscopy (EDS). The maximum absorbance of UV-vis spectra was at 432 nm. TEM analysis reveals that the shape of the most biosynthesized AgNPs was in spherical forms and the average of particle size was 18 nm. EDS analysis exhibits strong signals of silver element. In addition, green synthesized AgNPs show high and efficient antimicrobial activities against two food-borne pathogens (E. coli O157:H7 and L. monocytogenes). TEM and scanning electron microscopy images reveal that there were significant shrinkage and damage of bacterial cell wall, and the leakage or loss of bacterial intracellular contents. A significant reduction of bacterial counts just after 4 h of exposure was observed. These results indicate that green synthesized AgNPs can be utilized as an antimicrobial means to inhibit the growth of pathogenic bacteria for applications in agricultural and food industries. Outputs: <ol> <li>Alsammarraie, A.K., Wang, W., Zhou, P., Mustapha, A., Lin, M. 2018. Green synthesis of silver nanoparticles using turmeric extracts and investigation of their antibacterial activities. Colloids Surf. B Biointerfaces. 171, 398-405.</li> <li>Liu, L., Kerr, W.L., Kong, F., Dee, D.R., Lin, M. 2018. Influence of nano-fibrillated cellulose (NFC) on starch digestion and glucose absorption. Polym. 196, 146-153.</li> <li>Tian, K., Chen, X., Luan B., Singh, P., Yang, Z., Gates, K.S., Lin, M., Mustapha, A., Gu, L.-Q. 2018. Single LNA-enhanced genetic discrimination of foodborne pathogenic serotype in a nanopore. ACS nano. 12, 4194-4205.</li> <li>Alsammarraie, A.K., Lin, M., Mustapha, A., Lin, H., Chen, X., Chen, Y., Wang, H., Huang, M. 2018.  Rapid determination of thiabendazole in juice by SERS coupled with novel gold nanosubstrates. Food Chem. 259, 219-225. </li> </ol>   Station: University of Wisconsin-Madison PI: Sundaram Gunasekaran Investigators/Participants: Sundaram Gunasekaran and Joel Pedersen Objective(s) Addressed: # 1, 2, 3, & 4 Summary of Work: Nanomaterials synthesis and biosensor fabrication for evaluating various analytes to ensure food (e.g., deleterious microbiological and chemical constituents) and environmental (e.g., heavy metals, prions) safety. Sustainable development of nanotechnology via molecular-level understanding of the interaction of nanomaterials with biological interfaces, both to design applications that interface with biological systems and to evaluate the potential risks posed by release of nanoscale materials into the environment. Outputs: <ol> <li>Anu Prathap MU, Sadak O, Gunasekaran S. 2018. Rapid and scalable synthesis of ultrathin zeolitic imidazole framework (ZIF-8) and its use for the detection of trace levels of nitroaromatic explosives. Adv Sustainable Systems 2 (10),1800053</li> <li>Xu X, Guo Y, Wang L, He K, Guo Y, Wang XQ, Gunasekaran S. 2018. Hapten-grafted programmed probe as corecognition element for competitive immunosensor to detect acetamiprid residue in agricultural products. J Ag and Fd Chem 66(29):7815-7821.</li> <li>Sadak O, Sundramoorthy AK, Gunasekaran S. 2018. Facile and Green Synthesis of Highly Conductive Graphene. Carbon 138:108-117.</li> <li>Urena-Saborio H, Alfaro-Viquez E, Esquivel-Alvarado D, Madrigal-Carballo S, Gunasekaran S. 2018. Electrospun plant mucilage nanofibers as biocompatible scaffolds for cell proliferation. International J. of Biological Macromolecules 115:1218-1224.</li> <li>You YS, Lim S, Hahn J, Choi YJ, Gunasekaran S. 2018. Bifunctional linker-based immunosensing for rapid and visible detection of bacteria in real matrices. Biosens Bioelectron 100:389-95</li> <li>Xu XH, Guo YN, Wang XY, Li W, Qi PP, Wang Z, Gunasekaran S, Wang Q. 2018. Sensitive detection of pesticides by a highly luminescent metal-organic framework. Sensor Actuat B-Chem 260:339-45</li> <li>Wang YC, Mohan CO, Guan JH, Ravishankar CN, Gunasekaran S. 2018. Chitosan and gold nanoparticles-based thermal history indicators and frozen indicators for perishable and temperature-sensitive products. Food Control 85:186-93</li> <li>Melby, E. S.; Allen, C. R.; Foreman-Ortiz, I. U.; Caudill. E. R.; Kuech, T. R.; Vartanian, A. M.; Zhang, X.; Murphy, C. J.; Hernandez, R.; Pedersen, J. A. Peripheral membrane proteins dramatically alter nanoparticle interaction at lipid bilayer interfaces. Langmuir 2018, 34, 10793-10805.</li> <li>Mensch, A. C.; Buchman, J. T.; Haynes, C. L.; Pedersen, J. A.; Hamers, R. J. Quaternary amine-terminated quantum dots induce structural changes to supported lipid bilayers. Langmuir 2018, 34.</li> <li>Frank, B. P.; Durkin, D. P.; Caudill, E. R.; Zhu, L.; Curry, M. L.; Pedersen, J. A.; Fairbrother, D. H. Impact of silanization on the dispersion properties and biodegradability of nanocellulose. ACS Appl. Nano Mater. 2018, 1.</li> <li>Olenick, L. L.; Troiano, J. M.; Vartanian, A.; Melby, E. S.; Mensch, A. C.; Zhang, L.; Hong, J.; Mesele, O.; Qiu, T.; Bozich, J.; Lohse, S.; Zhang, X.; Kuech, T. R.; Millevolte, A.; Gunsolus, I.; McGeachy, A. C.; Doğangün, M.; Li, T.; Hu, D.; Walter, S. R.; Mohaimani, A.; Schmoldt, A.; Torelli, M. D.; Hurley, K. R.; Dalluge, J.; Chong, G.; Feng, Z. V.; Haynes, C. L.; Hamers, R. J.; Pedersen, J. A.; Cui, Q.; Hernandez, R.; Klaper, R.; Orr, G.; Murphy, C. J.; Geiger, F. M. Lipid corona formation from nanoparticle interactions with bilayers and membrane-specific biological outcomes. Chem 2018, 4, 2709-2723.</li> <li>Plummer, I. H.; Johnson, C. J.; Chesney, A. R.; Pedersen, J. A.; Samuel, M. D. Mineral licks as environmental reservoirs for chronic wasting disease prions. PLoS ONE 2018, 13, e0196745.</li> </ol>   Planned Activities: The team has identified the following subgroups with their associated participants to facilitate discussion and collaboration among the members. <ol> <li>Inline measurement processing <ol> <li>Chenxu Yu*</li> <li>Mingshi Lin</li> <li>Yanbin Li</li> <li>Ramaraja Ramasamy</li> <li>Paul Takhistov</li> </ol> </li> <li>Matrix challenge <ol> <li>Ramaraja Ramasamy</li> <li>Evangelyn Alocilja*</li> <li>Paul Takhistov</li> <li>Sundaram Gunasekaran</li> <li>Margaret Frey</li> </ol> </li> <li>False +/- (in general) <ol> <li>Ramaraja Ramasamy*</li> <li>Sundaram Gunasekaran</li> <li>Evangelyn Alocilja</li> </ol> </li> <li>Dead vs. Live cells <ol> <li>Ramaraja Ramasamy</li> <li>Evangelyn Alocilja</li> <li>Jeremy Tzeng*</li> <li>Olga Tsyusko</li> </ol> </li> </ol>

Publications

<ol> <li>Cao, L.L., Q. Zhang, H. Dai, Y.C. Fu, and Li. 2018. Separation/concentration-signal-amplification in-one method based on electrochemical conversion of magnetic nanoparticles for electrochemical biosensing. Electroanalysis 30(3):517-524. DOI: 10.1002/elan.201700653</li> <li>Dai, H., Y.Q. Li, Y.C. Fu, and Li. 2018. Enzyme catalysis induced polymer growth in nanochannels: A new approach to regulate ion transport and to study enzyme kinetics in nanospace. Electroanalysis 30(2):328-335 (available online Dec. 18, 2017). DOI:10.1002/elan.201700703</li> <li>Dai, H., Y.Q. Li, Q. Zhang, Y.C. Fu, and Li. 2018. A colorimetric biosensor based on enzyme-catalysis-induced production of inorganic nanoparticles for sensitive detection of glucose in white grape wine. RSC Advances 8:33960-33967. DOI: 10.1039/c8ra06347h</li> <li>Hu, Q.Q., R.H. Wang, H. Wang, M.F. Slavik and Li. 2018. Selection of acrylamide-specific aptamers by a quartz crystal microbalance combined SELEX method and their application in rapid and specific detection of acrylamide. Sensors and Actuators: B: Chemical 273:220-227. doi.org/10.1016/j.snb.2018.06.033</li> <li>Li, Z.S., G.S. Zhou, H. Dai, M.Y. Yang, Y.C. Fu, Y.B. Ying, and Li. 2018. Biomineralization-mimetic preparation of hybrid membranes with ultra-high load of pristine metal-organic frameworks grew on silk nanofibers for hazards collection in water. Journal of Materials Chemistry A 6(8):3402-3413 (published online on December 5, 2017). DOI:10.1039/C7TA06924C</li> <li>Wang, H., L.J. Wang, Q.Q. Hu, R.H. Wang, Li and M. Kidd. 2018. Rapid and sensitive detection of Campylobacter jejuni in poultry products using a nanoparticles-based piezoelectric immunosensor integrated with magnetic immunoseparation. Journal of Food Protection 81(8):1321-1330. doi:10.4315/0362-028X.JFP-17-381</li> <li>Wang, L.J., R.H. Wang, H. Wei, and Li. 2018. 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Impact Statements

The participants were invited to submit a full proposal entitled "Data-Driven Integrated Approach for Safe and Traceable Fresh Produce Supply Chains Across the Border" in responding to the USDA-NIFA-AFRI-006553 funding opportunity.

Date of Annual Report: 06/06/2019

Report Information

Annual Meeting Dates: 05/23/2019 - 05/25/2019
Period the Report Covers: 10/01/2018 - 09/30/2019

Participants

Paul Takhistov, Rutgers Univ. (host)
Vangie Alocilja, Michigan State University
S. Gunasekaran, UW Madison
Anhong Zhou, Utah State University
Olga Tsyusko, University of Kentucky
David Monks, NC State (acting as project administrator for Steve Lommel)
Jeremy Tzeng, Clemson University (chair)
Mengshi Lin, Univ. of Missouri (vice-chair)
Daniel Jenkins, Univ. of Hawaii (secretary)

Brief Summary of Minutes

Minutes Attachment:

2019 NC-1194 Annual Meeting Minutes.docx

Accomplishments

NC 1194 Annual Report Accomplishments 10/01/2018 – 09/30/2019 Station: Arizona (University of Arizona) PI: Dr. Jeong-Yeol Yoon Investigators/Participants: Soo Chung (PhD student), Robin E. Sweeney (PhD student), Katherine E. Klug (PhD student), Tiffany-Heather Ulep (PhD student), Benjamin Alouidor (MS student), Vina Nguyen (MS student), Lane E. Breshears (undergraduate student), Trinny Tat (undergraduate student), Christian M. Jennings (undergraduate student), Elizabeth Budiman (undergraduate student), Sean Perea (undergraduate student), Alexander S. Day (undergraduate student), Katelyn Sosnowski (undergraduate student), Alexa Shumaker (undergraduate student), Brandon T. Nguyen (undergraduate student), Raymond K. Wong (Professor; Collaborator in Medical Pharmacology), Lingling An (Professor; Collaborator in Biosystems Engineering and Statistics), Walter Q. Betancourt (Professor; Collaborator in Soil, Water and Environmental Science), and Kelly A. Reynolds (Professor; Collaborator in Public Health). Accomplishments: There is a growing need to develop a handheld, smartphone-based biosensor that can detect the type and concentration of pathogens from myriads of food (fresh produce and meat) and water (waste and irrigation) samples, as well as urine, blood, and tissue samples from animal and human subjects. These biosensors must be designed and manufactured to be easy-to-use, all-in-one, and extremely sensitive (down to single cell level, single genomic copy level, or picogram protein level). During the period from October 1st, 2018 to September 30th, 2019, we have published 7 journal papers. In addition, there are four journal papers and one text book accepted in this period (not listed here – will be reported in the next year’s report). Some of our work have received recognitions from professional societies, such as “Most Downloaded Journal Paper” from IEEE (listed #4 below) and “Highly Cited Journal Paper” from Royal Society of Chemistry (RSC) (Anal. Meth. 8: 6591-6601; published in late 2016 but the recognition was made in 2019). Most importantly, our work on single copy level detection of norovirus was selected one of only 17 papers (out of >12,000 papers) to be press-released by American Chemical Society (ACS) Fall 2019 National Meeting & Expo at San Diego Convention Center. There was also a press conference, broadcasted live on YouTube, and subsequently picked up by numerous news media, including NPR News from Washington Headquarter. Media coverage is summarized in #5 shown below. We are also translating our handheld biosensor technology into real-world practice, through collaborating with Tucson Water and Korea Institute of Ocean Science and Technology (KIOST), who have also provided research grants to our lab. Station: Hawaii (University of Hawaii) PI: Daniel M. Jenkins Investigators/Participants: Daniel M. Jenkins (Professor); Lena Diaz (PhD Student / Graduate Assistant) Accomplishments: Our work has focused on two main projects- the development of open-source hardware and software for a wireless potentiostat project to facilitate commercialization of nanotechnology-based sensors for field use, and development of molecular probes and classification algorithms for discrimination of single nucleotide polymorphisms (SNPs) in pathogenic organisms. The potentiostat project, “ABE-Stat” was developed in collaboration with project participants at the Universities of Florida and Georgia, and has been fully characterized and documented in an invited open-access issue of the Journal of the Electrochemical Society. The device is the first open-source project of it’s kind with a fully wireless smart-phone interface, and to incorporate Electrochemical Impedance Spectroscopy (EIS) capability across a wide spectrum (0.1 Hz to 100 kHz). We have shared prototypes of the device with numerous researchers in the US and abroad. A second iteration of the device was designed and assembled to improve signal to noise performance especially for dynamic voltammetry applications, and to address some issues in consistency of EIS measurements across the spectrum that resulted in discontinuities in recorded spectra in for non-linear systems using the original design. We are currently developing firmware updates to more fully characterize this system in anticipation of making this new design more widely available to the research community. For our molecular probe design we incorporated mismatched ribonucleotides into quenched fluorescence DNA probes (i.e. strands terminated with fluorophore and quencher molecules), and observed changes in fluorescence due to differential probe annealing and RNAase actuated cleavage to different SNP variants of the target gene. While individual probes exhibited some cross reactivity with their non-target variants, characteristic patterns of the real-time reactions could reliably be used to predict which gene variant was present, and mixtures of probes could be used to identify presence of variants of the gene even for heterozygous types. Station: Iowa (Iowa State University) PI: Chenxu Yu, Investigators/Participants: Chenxu Yu, Jonathan Claussen Accomplishments: Objective 1. Develop new technologies for characterizing fundamental nanoscale processes. In this period, we continued evaluating production and presence of carbon nanoparticles in foods, and their fluorescence and bioluminescence properties. It furthered our understanding of naturally occurring nanoscale processes in food matrix which may lead to better utilization of these nano-phenomena. Objective 2. Construct and characterize self-assembled nanostructures. We continued working on development of nano-vaccines using self-assembled nanostructures as carriers and cloned viral protein sigma 1 as a target recognition mechanism for improved delivery. Objective 3. Develop devices and systems incorporating microfabrication and nanotechnology. We continued our work on SERS imaging to achieve rapid detection of pathogens in low moisture foods. We also continued investigating the potential of using portable Raman imaging to diagnose Chronic Downing disease in deer. Station: Kentucky (University of Kentucky) PI: Olga Tsyusko (University of Kentucky, College of Agriculture Food and Environment, Department of Plant and Soil Sciences) Investigators/Participants: Olga Tsyusko: Ph.D. Students: Anye Wamucho, Stuart Lichtenberg, and Jarad Cochran; Collaborators: Evangelyn Alocilia (NC 1194 member), Jason Unrine, Isabel Escobar, Andrew Morris, Claus Svendsen, Carolin Shultz and David Spurgeon Accomplishments: The ongoing projects in my laboratory can be divided into four categories: Toxicity and mechanisms from exposure to pristine and environmentally transformed metal and metal oxide nanomaterial. Together with my Ph.D. students, Daniel Starnes and Anye Wamucho, we examined bioavailability, toxicity and its underlying mechanisms from exposure of a model organism soil nematode Caenorhabditis elegans, to silver and zinc oxide nanomaterials in their pristine and environmentally modified (aged) forms. Among transformed nanomaterials were silver sulfide, zinc phosphate and zinc sulfide. We were among the first to examine multigenerational effects from the exposure to silver nanomaterials. Our previous studies demonstrated that exposure to pristine silver nanomaterials induced reproductive sensitivity in as early as second generation, which persisted for nine generations without recovery, even when exposure has been stopped. We investigated whether genomic mutations or/and epigenetic modifications can explain such heightened sensitivity. While increase in germline mutations were observed after exposures to all silver treatments, changes in histone methylation (at H3K4me2 and H3K9me3) demonstrate stronger correlation with the reproductive toxicity. Efficiency and non-target effect of nanocomposites. In this research we examine efficiency and non-targeted toxicity of chitosan/dsRNA polyplex nanoparticles using C. elegans. We demonstrate that chitosan/dsRNA polyplex nanoparticles can knockdown targeted gene effectively than naked dsRNA. In addition,  we show that chitosan/dsRNA polyplex nanoparticles introduce dsRNA into cells through clathrin-mediated endocytosis, which is a different mechanism from canonical pathway via sid-1 and sid-2. Finally, our results suggest that  chitosan, as either polyplex nanoparticles or alone, downregulates the expression of myosin. Examining toxicity of phosphorene and its potential for degradation/removal of PFAS from drinking water.  The third project focuses on examining toxicity of 2D nanomaterial phosphorene on C. elegans with the purpose of developing “safe” phosphorene membranes. The overarching goal of this project is to develop novel nanocomposite membranes with minimal toxicity and to test their potential to remove per- and polyfluoroalkyl substances (PFAS). The specific aims are 1) to test in vivo toxicity of phosphorene nanomaterials in a free form and in filtrate after nanoparticles are embedded into the membrane on C. elegans and 2) relying on phosphorene’s photocatalytic properties, to degrade PFAS, that accumulate on the membrane surface, and examine toxicity of PFAS and their breakdown products to C. elegans. Testing efficiency of iron oxide nanomaterials against antimicrobial resistant (AMR) bacteria in C. elegans. This is in collaboration with NC 1194 member Dr. Evangelyn Alocilja.  My new PhD student, Jarad Cochran, focuses on testing toxicity of antibiotic resistant and non-resistant strain Klebsiella pneumoniae before and after treatment with iron oxide nanomaterials to C. elegans. Currently we are examining toxicity (mortality, reproduction and internal ROS production) of iron oxide nanomaterials to C. elegans. The objective is to identify concentration which will not induce toxicity responses at the examined endpoints in C. elegans. After that both strains of K. pneumoniae will be treated to the “safe” for C. elegans concentration of iron oxide nanoparticles and will be fed to C. elegans to examine efficiency of the iron oxide nanomaterials against the AMR K. pneumoniae. Station: Michigan (Michigan State University) PI: Dr. Evangelyn Alocilja, Professor, Biosystems and Agricultural Engineering Accomplishments: We develop magnetic and gold nanoparticles for bacterial extraction and detection in complex matrices for health, food safety, and water quality. We have optimized our newly synthesized magnetic nanoparticles that can be used to isolate and concentrate pathogenic bacteria from food, water, and clinical samples. We have optimized our dextrin-capped gold nanoparticles for colorimetric signal generation in genomic DNA detection. We are conducting studies to determine the concentration effect of our magnetic nanoparticles in human, foodborne, and waterborne pathogens. We are developing new antimicrobial nanoparticles to help reduce foodborne illness. I received a Balik Scientist Award from the Philippine government in 2018 which funded my travel to present my research as well as allow me to establish and strengthen new research collaborations. I received a travel fellowship from Ehime University which allowed me to travel to Japan and present my research as well as establish new research collaboration with faculty from Ehime University. We published 9 peer-reviewed journal papers and made 26 conference/meeting presentations. Station: Missouri (University of Missouri) PI: Mengshi Lin, Professor, Food Science Program, University of Missouri Investigators/Participants: Fouad Alsammarraie (PhD student), Lin Sun (PhD student) Accomplishments: Our objectives are to develop new technologies for characterizing fundamental nanoscale processes; construct and characterize self-assembled nanostructures; and develop devices and systems using nanotechnology. In this reporting period, we developed a nanocomposite based on nanofibrillar cellulose (NFC) integrated with gold-silver (core-shell) nanoparticles (Au@Ag NPs) as a novel surface-enhanced Raman spectroscopy (SERS) substrate. SERS performance of NFC/Au@Ag NPs nanocomposite was tested by 4-mercaptobenzoic acid (4-MBA). Cellulose nanofibril network was a suitable platform that allowed Au@Ag NPs to be evenly distributed and stabilized over the substrate, providing more SERS hotspots for the measurement. Two pesticides, thiram and paraquat, were successfully detected either individually or as a mixture in lettuce by the nanocomposite. Strong Raman scattering signals for both thiram and paraquat were obtained with the Raman intensities ~8 times higher than the ones that were acquired by NFC/Au NPs nanocomposite. Characteristic peaks were clearly observable in all SERS spectra even at a low concentration of 10 µg/L of pesticides. Limit of detection values of about 71 µg/L and 46 µg/L were obtained for thiram and paraquat, respectively. Satisfactory SERS performance, reproducibility and sensitivity of NFC/Au@Ag NPs nanocomposite validate its applicability for real-world analysis to monitor pesticides and other contaminants in complex food matrices within a short acquisition time. Station: New York (Cornell University) PI:  Margaret Frey Investigators/Participants: Soshana Smith, Katarina Goodge, Michael Delaney Accomplishments: We continue to work on producing submicron fibers with surface properties suitable to capture and concentrate target chemicals, pathogens or biological molecules from liquids.  Our latest efforts focus on attaching specific anti-bodies to nanofiber surfaces and measuring capture capabilities from fluids and in microfluidic channels. Two types of fibers are described below. Biotin Functionalized Silver-Doped Carbon Nanofibers for Selective Capture of E.coli in Microfluidic Systems: Silver-doped carbon nanofibers are used as the base of material for the selective capture of E.coli in microfluidic systems. As-spun PAN fiber, PAN based carbon nanofibers, as-spun PAN/AgNO3, Ag doped nanofibers,and Ag doped nanofibers coated with poly(3,4-ethylenedioxythiophene): polystyrenesulfonate  (PEDOT:PSS) were studied using various characterization techniques. Energy-dispersive X-ray spectroscopy was used to confirm the confirm the presence of certain elements in the fibers such as the sulfur in PEDOT:PSS. While 4 probe testing was used to study the conductivity of the various fibers. After various tests, it was determined that the PEDOT: PSS coating was not needed to ensure the success of the future steps. Therefore silver-doped carbon nanofibers were chosen as the desired substrate. Antibodies were immobilized on the surface of the silver-doped nanofibers via a 3-stepprocess. The negatively charged silver particles present on the surface of the nanofibers provide a suitable substrate for positively charged biotinylated poly(l-lysine)-g-poly(ethylene glycol) (PLL-g-PEG)FITC to attach. PLL-g-PEG FITC was initially used to get a visual conformation that the conjugate attachment worked using confocal microscopy. A similar conjugate, PLL-g-PEG biotin, was used for future experiments to take advantage of the strong affinity of biotin for streptavidin. After the addition of the conjugate, streptavidin and a biotinylated anti E coli antibody are then added system to selectively capture E. coli cells with high efficiency. We will demonstrate the fibers ability for isolation, detection, sequential collection of E coli. Functionalizing Cellulose-based Nanofibers Using “Click” Chemistry: This project aims to address the ongoing challenges of disease diagnostic technology by leveraging the high specific surface area and functionality of nanofibers to design highly sensitive and accurate bioassays. Specifically, the surface chemical modification of cellulose acetate nanofibers was studied to determine the most efficient and effective method to use as the intermediate step in attaching functional molecules via click chemistry. Cellulose acetate was electrospun into nanofibers that were chemically modified either with or without a deacetylation step. The as-spun and regenerated fibrous mats have alkyne moieties covalently attached to the nanofibers that selectively “click” with corresponding azide biotin conjugates. Two methods of alkyne-to-fiber attachment were tested and optimized to achieve practically useful degrees of substitution of the respective moieties on the repeat units. The Huisgen [3+2] copper-catalyzed azide-alkyne cycloaddition (CuAAC) “click” reaction was used to model the potential of click degree of substitution by using biotin as a model biomolecule and streptavidin-FITC as a fluorescence marker. FTIR and Raman were used as initial characterization techniques to confirm successful reaction products. EDX was used to map the biotin clicked onto the mats as well as confocal microscopy to confirm the distribution and degree of substitution of the biotin conjugate. Station: South Carolina (Clemson University) PI: Dr. Tzuen-Rong Jeremy Tzeng Accomplishments: Pathogen attachment is a complex phenomenon and vital process for successful initiation of infection in the host. Bacterial pathogens utilize two primary mechanisms to adhere onto host cells, namely carbohydrate-protein recognition and protein-protein interaction. The adhesion structures have a high degree of preference for a particular host-cell receptor. We have developed nanoparticles functionalized with specific receptors and evaluated their ability for selective binding and killing of pathogens. During this report period, we have developed iron-oxide nanoparticles functionalized with carbohydrates specific for Neisseria gonorrhoeae and conducted both in vitro and in vivo mouse models. In addition, we have developed and evaluated X-ray excited luminescent chemical imaging (XELCI) for non-invasive imaging of implant related infections. Station: South Dakota (South Dakota State University) PI: Zhengrong Gu Investigators/Participants: Undergraduates: Nelson, Zebadiah Peter; Emily Leupp; VanWell, Elliot; Held, Logan; PhD students: Hummel, Matthew, Shun Lu. Accomplishments: We developed new technologies for characterizing fundamental nanoscale processes. We developed electrochemical analysis, including impedance, differential pulse voltammetry of biomolecule interactions such as DNA chains hybridization, antibody-antigens interaction in nano-scale. We constructed and characterized self-assembled nanostructures. Nickel oxide nanoparticles anchored on three-dimensional (3D) carbonized eggshell membrane (NiO/C) are synthesized by a green hydrothermal approach followed by a pyrolysis in the nitrogen atmosphere at 500 ℃. The introduction of eggshell membrane (ESM) provides a microreactor for the formation of Ni(OH)2/ESM in which Ni(OH)2 is formed by adsorption of ESM during hydrothermal process, which not only exposes more catalytic activity edge but also transfers biowaste into useful nanomaterials. The carbonized ESM exhibited its robust and porous structure after heat treatment, which provided a stable support and electronic transmission channels that endowing the Ni2+ to Ni3+ pre-oxidation process to boost urea electrooxidation and detection. Various physical characterization methods were applied to confirm the structure, consistence and properties of the as-synthesized sample.We also prepared kappa-carrageenan (KC) and hierarchical porous activated carbon (HPAC) auto-assembled nano-composite. The alkali lignin derivate HPAC was synthesized with a confidential atmospheric plasma processes. Due to KC’s unique interlocking helix-structure, the KC-HPAC structured composite demonstrated high surface area and highly porous structure, in which the KC helical strands extend from the surface of HPAC. TEM images also demonstrated both large, irregular dispersed pores of >250nm and nano-sized, regularly interspersed pores in the prepared HPAC and KC-HPAC composite. The hierarchical pore structure of HPAC is attributed to the rapid conversion in atmospheric plasma process. KC’s pores are a result of the spaces left between the interlocking helices of the linear disaccharide, which auto-assembles to this conformation after dissolving in water and subsequently constituting as a gel. We developed devices and systems incorporating microfabrication and nanotechnology. The electrocatalytic performance of NiO/C electrode toward urea oxidation and urea detection in alkaline solution was evaluated using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), linear sweep voltammetry (LSV), square wave voltammetry (SWV) and chronoamperometry (CA). Results show that the as-prepared electrode possesses an onset oxidation potential of 1.47 V (vs. RHE) and a peak current density of 1.86 mA/cm2 in 1.0 M KOH and 0.33 M urea solutions. Moreover, NiO/C modified electrode exhibits an outstanding performance for urea determination with two linear ranges of 0.05-2.5 mM, a limit of detection (LOD) of &sim;20 μM, and fast response time (within 7 s). Furthermore, the stability and anti-interference performance of the developed electrochemical sensor was also measured. The good selectivity of the electrochemical urea sensor in a diluted urea sample containing different interfering species, such as glucose (Glu), Na+ ions, K+ ions, and uric acid (UA) were verified with stairs response towards urea of the current electrode when successive added urea, while insignificant responses to the other interfering species. Significantly, this work offered a green method for fabricating 3D nanostructure by using a biowaste ESM as template, setting up a typical example for producing new value-added nanomaterials with urea electrooxidation. Multiple HPAC-KC composite were tested as electrode for detecting dopamine in solution. It was hypothesized that due to HPAC’s higher conductivity than KC, the limit of detection would be higher with an increasing proportion of HPAC. This hypothesis was confirmed. As predicted, having a higher proportion of conductive HPAC would both provide a higher amplitude of peak current and more sites for KC helices to adsorb physically. As a result, the limit of detection with a S/N ratio of 3 was highest for 2500:1 HPAC: KC at 0.4 umol/L and a linear range of 1 umol /L– 600 umol/L. Interestingly, the limit of detection did not scale uniformly with the increasing concentration of HPAC. Instead, the second highest limit of detection was determined to be 100:1 HPAC: KC. The lowest limit of detection was encountered with the 500:1 HPAC: KC film. One potential explanation for this phenomenon could lie in the orientation of the KC helices on the HPAC’s surface. KC’s helices best film forming occurs in 1.0-2.5% solutions by mass in 1% acetic acid. In the 200:1-1250:1 HPAC: KC range, this interaction between acetic acid and KC is disrupted due to the HPAC’s own interactions with KC and acetic acid. By increasing the ratio, HPAC’s interactions with KC dominate, while lower ratios allow KC to interact with acetic acid and form more stable, protonated gels, which also benefit detection of dopamine. Furthermore, the innovative HPAC-KC platform is competitive with other reported biosensors of dopamine in terms of linear range and limit of detection. A major advantage the HPAC: KC film has over the majority of dopamine sensors is its simple method of preparation and low-cost materials. Notably, no adhesive polymer such as PTFE, Nafion, or PVDF was necessary in adhering the film to the surface of the glass carbon electrode. Instead, the adhesive properties of the composite itself were relied upon with a high degree of success. Station: Utah (Utah State University) PI: Anhong Zhou Investigators/Participants: Graduate students: Han Zhang, Wei Zhang; Undergraduates: Reem Ghabayen, Kamila Khamidova Accomplishments: Understanding of the DNA-mediated electron transfer characteristics on sensor surface plays essential roles in design of highly sensitive DNA biosensors. We have applied a variety of electrochemical techniques, including cyclic voltammetry, differential pulse voltammetry, and electrochemical impedance spectroscopy to quantitatively assess the electron transfer behaviors of each step of construction of DNA biosensors, such as probe DNA immobilization, probe-target hybridized surface, capture probe DNA hybridization, and electroactive ALP-linked enzyme. We obtained the optimal sensor design based on these assessment results. Starting with the first step of immobilizing probe DNA (thiolated oligonucleotides), the construction of DNA biosensor includes multiple self-assembled steps to finally form highly specific DNA structure on sensor surface for recognizing the target DNA sequence. The experimental conditions of each self-assembled step are optimized. The overall performance such as sensitivity, selectivity, detection limit, and linear range have been evaluated for hybridization with complementary, single-base, three-base, and five-base mismatched target sequences. A prototype microfluidic device was fabricated using photolithograph techniques, and tested in detection of various target sequences (with different base-mismatch) to achieve satisfactory results. Our DNA biosensor for the first time realized the separation of the probe/target DNA immobilized sensor part (disposable) from the detection of electron-transfer sensor part (reusable), making it well suitable for multiplex detection of other species-specific target DNA with similar sequences (to test specificity). In addition, to address the Objective#3, we initiated a collaboration with Prof. Heloisa Rutigliano’s group from USU Animal Science Department in using non-invasive Raman spectroscopy technique to characterize the nanoscale extracellular vesicles (EVs) isolated from different animal cell samples. Further, we also designed and fabricated a prototype of smartphone based colorimetric biosensor for the detection of blood glucose. We have tried it successfully with artificial urine samples, and we are testing this biosensor for detecting blood glucose in cows at different pregnancy stages. In this past year, we have been successfully testing our DNA sensor with the target sequence specific to human genotype of Cryptosporidium. Our results indicate a detection of limit of target sequence with six times lower sensitivity, compared to the literature reported method. Station: Virginia (Virginia Polytechnic Institute and State University) PI: Chenming (Mike) Zhang Investigators/Participants: Yi Lu (graduate student) Accomplishments: We have constructed a chimeric protein based on Hepatitis B core antigen.  The chimeric protein is expressed in E. coli and purified by various chromatographic techniques.  The purified protein could self-assembled into virus like particles, which was characterized by scanning electron microscopy.  The virus-like particle is studied as a potential vaccine against a swine disease. Planned Activities: The NC-1194 members are working on a review article about antimicrobial resistance (AMR) that is a global public health challenge requiring a global multi-disciplinary approach.  Due to globalization, AMR anywhere is a threat to health everywhere. New antibiotic resistance mechanisms are spreading globally, threatening our ability to treat common infectious diseases, resulting in prolonged illness, disability, and death. In addition to the toll on human life, antibiotic-resistant infections add considerable avoidable costs to the already overburdened US health care system. Studies have estimated that, in the US, antibiotic resistance adds $20 billion in excess direct health care costs, with additional costs to society for lost productivity as high as $35 billion a year. Globally, failure to act on drug-resistant infections will lead to 10 million extra deaths a year and cost the global economy $100 trillion by 2050. The loss of effective antibiotic treatments will also undermine treatment of infectious complications in patients with other diseases. Many medical advances, such as joint replacements, organ transplants, cancer therapy, and others, are dependent on the ability to fight infections with antibiotics. If the ability to effectively treat those infections is lost, the ability to safely offer people the life-saving and life-improving modern medical advances will be lost with it. Thus, this paper will present scientific and engineering challenges of AMR in the human population, food and animal production, and the environment. The convergent multidisciplinary knowledge generated will be designed to stimulate new inquiries and challenge new discoveries toward AMR reduction. The members who have expressed interest in this review article include: Evangelyn Alocilja, Olga Tsyusko, Jeremy Tzeng,  Paul Takhistov,  Chenxu Yu, Zhengrong Gu, Mengshi Lin, Carmen Gomes, Jonathan Claussen, Jeong-Yeol Yoon, Anhong Zhou, Yi-Cheng Wang, Eric McLamore, Yanbin Li,  Elizabeth Carraay.

Publications

Publications Chung, S., Breshears, L.E., Yoon, J.-Y. *, Smartphone Near Infrared Monitoring of Plant Stress, Computers and Electronics in Agriculture, 2018, 154: 93-98. Alouidor, B., Sweeney, R.E., Tat, T., Wong, R.K.* and Yoon, J.-Y. *, Microfluidic Point-of-care Ecarin Based Clotting and Chromogenic Assays for Monitoring Direct Thrombin Inhibitors, Journal of ExtraCorporeal Technology, 2019, 51: 29-37. Klug, K.E., Jennings, C.M., Lytal, N., An, L., and Yoon, J.-Y. *, Mie Scattering and Microparticle Based Characterization of Heavy Metal Ions and Classification by Statistical Inference Methods, Royal Society Open Science, 2019, 6: 190001. Sweeney, R.E., Nguyen, V., Alouidor, B., Budiman, E., Wong, R.K., and Yoon, J.-Y. *, Flow Rate and Raspberry Pi-based Paper Microfluidic Blood Coagulation Assay Device, IEEE Sensors Journal, 2019, 19(13): 4743-4751. Top 25 Most Downloaded IEEE Sensors Journal Papers in June 2019. Chung, S., Breshears, L.E., Perea, S., Morrison, C.M., Betancourt, W.Q., Reynolds, K.A., and Yoon, J.-Y. *, Smartphone-based Paper Microfluidic Particulometry of Norovirus from Environmental Water Samples at Single Copy Level, ACS Omega, 2019, 4(6): 11180-11188. Highlighted in ACS News Release and more. Tiffany-Heather Ulep, Alexander S. Day, Katelyn Sosnowski, Alexa Shumaker and Jeong-Yeol Yoon*, Interfacial Effect-based Quantification of Droplet Isothermal Nucleic Acid Amplification for Bacterial Infection, Scientific Reports, 2019, 9: 9629. Matthew V. Bills, Brandon T. Nguyen and Jeong-Yeol Yoon*, Simplified White Blood Cell Differential: An Inexpensive, Smartphone- and Paper-Based Blood Cell Count, IEEE Sensors Journal, 2019, 19(18): 7822-7828. M. Jenkins, B. E. Lee, S. Jun, J. Reyes-De-Corcuera, and E. S. McLamore, J. Electrochem. Soc., 166, B3056–B3065 (2019). Lv J, Yang Z, Xu W, Li S, Liang H, Ji C, Yu C, Zhu B, Lin X, Relationships between bacterial community and metabolites of sour meat at different temperature during the fermentation, Int J Food Microbiol. DOI: 10.1016/j.ijfoodmicro.2019.108286, 2019. Xiong X, He BY, Jiang D, Dong XF, Yu C and Qi H, Postmortem biochemical and textural changes in the sea cucumber Stichopus japonicus body wall (SJBW) during iced storage, LWT-Food Sci. Technol. DOI: 10.1016/j.lwt.2019.108705, 2019. Jiang D, Bai Y, He BY, Sui Y, Dong XF, Yu C and Qi H, Improvement of gel properties of mackerel mince by phlorotannin extracts from sporophyll of Undaria pinnatifidai and UVA induced cross-linking, J. Textural Studies, DOI:10.1111/jtxs.12480, 2019. Liu XY, Wang ZX, Zhang J, Song L, Li DY, Wu ZX, Zhu BW, Nakamura Y, Shahidi F, Yu C, and Zhou DY, Isolation and identification of zinc-chelating peptides from sea cucumber (Stichopus japonicus) protein hydrolysate, J. the Science of Food and Agriculture, DOI:10.1002/jsfa.9919, 2019. Liu XY, Wang ZX, Yin FW, Liu YX, Qin NB, Nakamura Y, Shahidi F, Yu C, Zhou DY, Zhu BW, Zinc-Chelating Mechanism of Sea Cucumber (Stichopus japonicus)-Derived Synthetic Peptides, Marine Drugs, 17, 438, DOI:10.3390/md17080438, 2019. Dong XF, Bai Y, Xu Z, Shi YX, Sun YH, Janaswamy S, Yu C, Qi H, Phlorotannins from Undaria pinnatifida Sporophyll: Extraction, Antioxidant, and Anti-Inflammatory Activities, Marine Drugs, 17, 434, DOI:10.3390/md17080434, 2019. Xiong X, He BY, Jiang D, Koosis A, Yu C, Qi H, Postmortem biochemical and textural changes in the Patinopecten yessoensis adductor muscle (PYAM) during iced storage, Inter. J. Food Properties, 22, 1024-1034, 2019. Sun H, Li DM, Jiang D, Dong XF, Yu C, Qi H, Protective polysaccharide extracts from sporophyll of Undaria pinnatifida to improve cookie quality, Food Measurement and Characterization, 13(1), 764-774, 2019. Zhang XY, Jiang D, Li DM, Yu C, Dong XF, Qi H, Characterization of a seafood-flavoring enzymatic hydrolysate from brown alga Laminaria japonica, Journal of Food Measurement and Characterization, 13(2), 1185-1194, 2019. Li JQ, Cao L, Li DM, Yu C, Tan MQ, Carbon dots from roasted mackerel (scomberomorus niphonius) for free radical scavenging, LWT-Food Sci. Technol., 111, 588-593, 2019. Bao R, Gao N, Lv J, Ji CF, Liang HP, Li SJ, Yu C, Wang ZY, Lin XP, Enhancement of Torularhodin Production in Rhodosporidium toruloides by Agrobacterium tumefaciens-Mediated Transformation and Culture Condition Optimization, Journal of Agricultural and Food Chemistry 67 (4), 1156-1164, 2019. Zhang, J, Hu J, Wang S, Lin X, Liang H, Li S, Yu C, Dong X, Ji C, Developing and Validating a UPLC-MS Method with a StageTip-Based Extraction for the Biogenic Amines Analysis in Fish, J Food Sci. 84(5),,1138-1144. doi: 10.1111/1750-3841, 2019. Li, SJ, Yu C, Pan JF, Ma RC, Lin XP and Dong XP, Combined effects of aging and low temperature, long time heating on pork toughness, Meat Science, 150, 33-39. 2019. Dong XP, Liu WT, Song X, Lin XY, Yu D, Yu C and Zhu BW, Characterization of Heat-Induced Water Adsorption of Sea Cucumber Body Wall, J. Food Science, 84(1), 92-100, 2019. Hu J, Zhao TF, Li SJ, Wang ZY, Wen CR, Wang HT, Yu C, Ji CF, Stability, microstructure, and digestibility of whey protein isolate – Tremella fuciformis polysaccharide complexes, Food Hydrocolloids, 89, 379-385, 2019. Mammadova N, Kokemuller R, Summers C, He Q, Ding S, Baron T, Yu C, Valentine R, Sakaguchi D, Kanthasamy A, Greenlee J and Greenlee MHW, Accelerated accumulation of retinal α-synuclein (pSer129) and tau, neuroinflammation and autophagic dysregulation in a seeded mouse model of Parkinson’s disease, Neurobiology of Disease, 121, 1-16, 2019. Wamucho, A., Unrine, J. M., Kieran, T. J., Glenn, T. C., Schultz, C. L., Farman, M. L., Svendsen, C., Spurgeon, D. J.,  Tsyusko, O. V. (2019). Genomic mutations after multigenerational exposure of Caenorhabditis elegans to pristine and sulfidized silver nanoparticles. Environmental Pollution vol. 254, pp. 113078 Li, J., Rodrigues, S., Tsyusko, O. V., Unrine, J. M. (2019). Comparing plant-insect trophic transfer of Cu from lab-synthesized nano-Cu(OH)(2) with a commercial nano-Cu(OH)(2) fungicide formulation. Environmental Chemistry vol. 16, pp. 411-418. Lichtenberg, S. S., Tsyusko, O. V., Palli, S. R., Unrine, J. M. (2019). Uptake and Bioactivity of Chitosan/Double-Stranded RNA Polyplex Nanoparticles in Caenorhabditis elegans. Environmental science & Technology  vol. 53, pp. 3832-3840. Starnes, D., Unrine, J. M., Chen, C., + Lichtenberg, S., Starnes, C., Svendsen, C., Kille, P., Morgan, J. S., # Baddar, Z. E., Spear, A., Bertsch, P. M., Chen, K. C., * Tsyusko, O. V. (2019). Toxicogenomic responses of Caenorhabditis elegans to pristine and transformed zinc oxide nanoparticles. Environmental Pollution vol. 247, pp. 917-926. Wamucho A., Heffley A.,  and Tsyusko* O.V. In Review. Multigenerational exposure of Caenorhabditis elegans to silver nanoparticles induces histone methylation changes. Submitted to Environmental Toxicology & Chemistry. Gordillo CM, Gomez AV, Sanchez HP, Pryg K, Shinners J, Murray N, Munoz-SG, Bencomo AA, Gomez, AB, Janapa LG, Enriquez NR, Martin M, Romero NS, and Alocilja EC. 2018. Magnetic Nanoparticle-based Biosensing Asasy Quantitatively Enhances Acid-Fast Bacilli Count in Paucibacillary Pulmonary Tuberculosis. Biosensors, 8(4):128-141. Alocilja EC, Sharief SA, Kriti N, and Chahal P. 2018. Combining Blockchain, DNA, and Nanotechnology for Product Authentication and Anti-Counterfeiting. Brand Protection Professional, (Dec. 20, 2018). Matta LL and Alocilja EC. 2018. Carbohydrate Ligands on Magnetic Nanoparticles for Centrifuge-free Extraction ofPathogenic Contaminants in Pasteurized Milk. J Food Protection, 81(12):1941-1949. (Dec. 2018) Zeeshan N, Daya KS, Tirumalai PS, and Alocilja E. 2018. Impedance and Magnetohydrodynamic Measurements for Label Free Detection and Differentiation of E. coli and S. aureus using Magnetic Nanoparticles. IEEE Transactions on NanoBioscience, 17(4):443-448. (Oct. 2018). Matta LL, Karuppuswami S, Chahal P, and Alocilja EC. 2018. AuNP-RF Sensor: An innovative application of RF technology for sensing pathogens electrically in liquids (SPEL) within the food supply chain. Biosensors and Bioelectronics, 111:152-158. Matta LL, Harrison J, Deol G, and Alocilja EC, 2018. Carbohydrate-functionalized Nano-Biosensor for Rapid Extraction of Pathogenic Bacteria Directly from Complex Liquids with Quick Detection Using Cyclic Voltammetry. IEEE Transactions on Nanotechnology, 17(5):1006-1013. Matta LL and Alocilja EC. 2018. Emerging nano-biosensing with suspended MNP microbial extraction and EANP labeling. Biosensors and Bioelectronics, 117:781-793 Sun, L., Yu, Z., Lin, M.* 2019. Synthesis of polyhedral gold nanostars as surface-enhanced Raman spectroscopy substrates for measurement of thiram in peach juice. Analyst. 144, 4820-4825. Yu, Z., Dhital, R., Wang, W., Sun, L., Zeng, W., Mustapha, A.*, Lin, M.* 2019. Development of multifunctional nanocomposites containing cellulose nanofibrils and soy proteins as food packaging material. Food Packaging and Shelf Life. 21, 100366. Asgari, S., Saberi, A.H., McClements, D.J., Lin, M.* 2019. Microemulsions as nanoreactors for synthesis of biopolymer nanoparticles. Trends Food Sci. Technol. 86, 118-130. Yu, Z., Wang, W., Dhital, R., Kong, F., Mustapha, M., Lin, M.* 2019. Antimicrobial effect and toxicity of cellulose nanofibril/silver nanoparticle nanocomposite prepared by an ultraviolet irradiation method. Colloids Surf. B. 180, 212-220. Yu, Z., Wang, W., Kong, F., Lin, M.*, Mustapha, A.* 2019. Cellulose nanofibril/silver nanoparticle composite as an active food packaging system and its toxicity to human colon cells. Int. J. Biol. Macromol. 129, 887-894. An oral presentation and a poster presentation were made at the International Fiber Society meeting in Austin, TX, October 27-30, 2019.  Manuscripts are being prepared on both projects and will be submitted for publication early in 2020. An implanted pH sensor read using radiography. Md. Arifuzzaman,  Paul W. Millhouse,  Yash Raval*,  Thomas B. Pace,  Caleb J. Behrend,  Shayesteh Beladi Behbahani*,  John D. DesJardins,  Tzuen-Rong J. Tzeng  and  Jeffrey N. Anker. Analyst, 2019, April 23; 144 (9): 2984-2993 Lu S., Hummel M., Gu Y., Gu, Z*, Trash to treasure: A novel chemical route to synthesis of NiO/C for hydrogen production. 2019. Intern. J. Hydrogen Energy 44 (31), 16144-16153. Yan Gu, Matthew Hummel, Zhengrong Gu*, Kasiviswanathan Muthukumarappan, Zhendong Zhao, 2019, Synthesis and Characterization of Allyl Terpene Maleate Monomers, submitted to Scientific Reports in 2019 July, under reviewing after revision in 2019 Oct. Shun Lu; Zhengrong Gu*; Xiaoteng Liu; Matthew Hummel; Yue Zhou; Keliang Wang; Yucheng Wang, 2019, A high-performance nickel oxide on carbonized eggshell membrane catalyst for electrocatalytic urea oxidation and detection, submitted to Applied Catalysis B:  Environmental 2019 August, under reviewing. Shun Lu; Zhengrong Gu*; Xiaoteng Liu; Matthew Hummel; 2019, Synthesis of Au@ZIF-8 nanoparticles for enhanced electrochemical detection of dopamine, submitted to the Journal of The Electrochemical Society 2019 Oct., under reviewing. Matthew Hummel, Shun Lu, Nelson, Zebadiah Peter, Zhengrong Gu*, 2019, Graphene-biopolymer composite electrode for detecting dopamine, submitted to Biosensors and Bioelectronics in 2019 August, under reviewing. Matthew Hummel, Shun Lu, Zhengrong Gu*, Erin Lee, Emily Leupp, 2019, High energy room temperature synthesis graphene from lignin – application in supercapacitor, submitted to Journal of Power Sources in 2019 August, under reviewing. Hoda ilkhani, Han Zhang, Anhong Zhou, “A novel three-dimensional microTAS chip for ultra-selective single base mismatched Cryptosporidium DNA biosensor”, Sensors & Actuators: B. Chemical, 2019, 282: 675–683. Han Zhang, Ethan Smith, Wei Zhang, Anhong Zhou, “Inkjet printed microfluidic paper-based analytical device (uPAD) for glucose colorimetric detection in artificial urine”, Biomedical Microdevices, 2019, 21:48. Han Zhang, Ana Caroline Silva, Wei Zhang, Heloisa Rutigliano, Anhong Zhou, “Raman spectroscopy characterization of extracellular vesicles derived from bovine placenta and peripheral blood mononuclear cells”, Analyst, 2019 (revision submitted).

Impact Statements

Station: Arizona (University of Arizona) PI: Dr. Jeong-Yeol Yoon Impacts: Economic Value or Efficiency: Alternative methods and devices have been proposed and successfully demonstrated for smartphone-based paper microfluidic devices. Specifically, rather than collecting overall light intensities (scattering or fluorescence) from paper microfluidic chips, we have evaluated the real-time flow distance (and subsequently the flow rate profile) and related it to the target presence and its concentration. This method provides assay reproducibility and is not affected by ambient lighting conditions. Economic Value or Efficiency: Another alternative method and device is the use of a smartphone-based fluorescence microscope to paper microfluidic devices, where the fluorescent particles were imaged and counted one-by-one. Through developing an original code, these counts were successfully related to the target presence and its concentration. This method provides extremely low limit of detection, down to a single copy virus level or to sub-femtogram (fg) scale. Environmental Quality: A large number of biosensor readings were collected and the assays were further perfected (improved accuracy and reproducibility) through using various machine learning algorithms. Environmental Quality: These demonstrations can significantly improve the applicability of our biosensor methods and devices, which can be used by a lay person in myriads of environmental conditions. Such demonstrations will protect the general public from potential health risks from food, water, and environment. Station: Hawaii (University of Hawaii) PI: Daniel M. Jenkins Impacts: We anticipate that our work will help facilitate the successful commercialization of nanotechnology based sensors for field detection of pathogens and contaminants of concern in agricultural and environmental systems, as our potentiostat design is highly portable (palm sized), affordable (bill of materials cost <$150), powerful (can easily be used for any standard electrochemical analysis with an intuitive wireless interface), and inherently can connect collected information to the internet for improved decision support. We also anticipate that our work on molecular probes and related platform developments can be used to improve the quality of information yielded in portable diagnostic platforms, including including incorporation of proper controls / quantitation, and extended ability to detect multiple genes of interest in a sample with no additional manipulation by the user. In our experience the largest barrier to adoption of new technologies for field use in agricultural and environmental applications is the relatively low yield of high quality information relative to required user input, so we believe that any incremental step to address this problem will help make these tools more accepted for efficient decision support in these systems. Station: Kentucky (University of Kentucky) PI: Olga Tsyusko Impacts: Results from these studies allows to differentiate between toxicity induced by released dissolved metal ions versus particulate effects. Out studies examine environmentally realistic scenarios where organisms are exposed to transformed form of nanomaterials, such as in wastewater, sludge and soils. This scenario is critical to take into account when predicting risk from the exposure to nanomaterials. We are also among the first to examine genomic and epigenetic multigenerational toxicity mechanisms of silver nanomaterials. Multigenerational toxicity can eventually result in the adverse effects on population and community levels and should be considered when evealuating ecological impact of the exposure to populations. The dsRNA nanocomposites, if properly developed, can be eventually applied in agriculture as “safe” nanopesticides. The identification of alternative pathway (endocytosis) for dsRNA can have implications for not only target but also nontarget species and should be considered when developing nanopesticides. However further improvements of in the delivery of dsRNA using nanocarriers can result in nanopesticide which will allow to reduce the total mass of dsRNA required currently for crop pest control. The phopsphorene studies may lead to groundbreaking discovery in treatment of drinking water against PFAS. Antibiotic resistant K. pneumoniae is responsible for hospital-associated infections in more susceptible elder and immunocompromised patients. There is a potential for iron oxide nanomaterials to be used when developing novel therapies against antimicrobial resistant K. pneumoniae and possibly other AMR bacteria. Station: Michigan (Michigan State University) PI: Dr. Evangelyn Alocilja, Professor, Biosystems and Agricultural Engineering Impacts: We made major progress in the development of rapid biosensing assays for drug-resistant pathogens that affect human health, food safety, and water quality. Our research on nanoparticle-based biosensors was also featured in a podcast by the National Nanotechnology Initiative of the US government. It is now a permanent resource online available to all those interested on the topic. Furthermore, our research was featured by the MSU Honors College brochure, which are distributed to new faculty and students at Michigan State University. In the international arena, we strengthened our Global Alliance for Rapid Diagnostics (GARD) by establishing centers of excellence (COEs) with our international collaborators. These COEs and alliances will provide the research infrastructure and networks for coordinating our global research in Latin America, Southeast Asia, and South Asia. We have also trained 10 undergraduate students, 2 PhD students, 1 MS student, and 1 high school teacher. We have also trained one scientist from the Philippines, one scientist from Nepal, and two scientists from India on the use of our technologies. These students and scientists will become the future research leaders in the emerging field of nanotechnology and biosensors. Station: Missouri (University of Missouri) PI: Mengshi Lin, Professor, Food Science Program, University of Missouri Impacts: Our study established an easy and cost-effective way to fabricate high-performance substrates for surface enhanced Raman spectroscopy. The method was able to detect trace amount of pesticides in juice products and fresh produce. The detection limits meet the maximum residue limits of EPA. The substrates generate high and reproducible Raman signals for pesticides. This substrate has great potential to be applied for the rapid measurements of chemical contaminants in foods. In addition, this project has provided training for two doctoral students and two Master's students. We have disseminated the results to the industry and scientific communities at professional conferences such as IFT, ACS, and IAFP. Station: New York (Cornell University) PI: Margaret Frey Impacts: One MS student will complete her thesis based on these projects. Additionally, one post-doctoral associate has been trained on new techniques based on this project and is currently applying for professional positions. Station: South Dakota (South Dakota State University) PI: Zhengrong Gu Impacts: Additional funding obtained based on preliminary data from the hatch project: Graphene based Electrochemical Sensor for Detection of Salmonella in Beed Products (SD Beef council, $72,658); South Dakota Biofilm Science and Engineering Center (NSF EpsCor Track I. $1.32M). Station: Utah (Utah State University) PI: Anhong Zhou Impacts: The biosensors devices developed by our research laboratory will benefit the water quality monitoring (DNA biosensor for Cryptosporidium DNA) and animal blood glucose detection (smartphone-based colorimetric glucose detection) as well as the discovery of new biomarker for monitoring of animal pregnancy stages (optical sensor for extracellular vesicles detection). These biosensor technologies will be found invaluable in environmental quality monitoring and agriculture biological systems (e.g., animal reproduction). Station: Virginia (Virginia Polytechnic Institute and State University) PI: Chenming (Mike) Zhang Impacts: PEDV is highly transmissible in pigs and causes diarrhea and vomiting, and death of 50-100 percent of infected piglets. Vaccination is the most effective method of preventing infectious diseases. Currently, killed-virus and modified-live vaccines are used clinically to control PED. However, both types of vaccines have inherent drawbacks. The goal of this work is to develop self-assemble protein nanoparticles as potential vaccines that carry immunogenic epitopes of PEDV. When inoculated in pigs, we anticipate the vaccine will elicit antibodies against PEDV to alleviate the clinical symptoms caused by PEDV and reduce the mortality in young piglets.

Date of Annual Report: 07/24/2020

Report Information

Annual Meeting Dates: 05/20/2020 - 05/21/2020
Period the Report Covers: 10/01/2019 - 09/30/2020

Participants

Yoon, Jeong-Yeol (jyyoon@arizona.edu) - University of Arizona; Li, Yanbin (yanbinli@uark.edu) - University of Arkansas; McLamore, Eric (emclamore@ufl.edu) - University of Florida; Reyes-de-Corcuera, Jose (jireyes@uga.edu) - University of Georgia; Jenkins, Daniel (danielje@hawaii.edu) - University of Hawaii; Gomes, Carmen (carmen@iastate.edu) - Iowa State University; Yu, Chenxu (chenxuyu@iastate.edu) – Iowa State University; Tsyusko, Olga (olga.tsyusko@uky.edu) – University of Kentucky; Cochran, Jarad (jaradcochran@uky.edu) – University of Kentucky; Alocilja, Evangelyn (alocilja@msu.edu) – Michigan State University, Lin, Mengshi (linme@missouri.edu) – University of Missouri; Takhistov, Paul (ptakhist@sebs.rutgers.edu) - Rutgers University; Mao, Yu (yu.mao@okstate.edu) – Oklahoma State University; Tzeng, Tzuen-Rong (TZUENRT@clemson.edu) – Clemson University; Zhou, Anhong (Anhong.Zhou@usu.edu) – Utah State University; Zhang, Chenming (cmzhang@vt.edu) – Virginia Polytechnic Institute; Gunasekaran, Sundaram (guna@wisc.edu) University of Wisconsin Madison; Lommel, Steve (slommel@ncsu.edu) – North Carolina State University / project administrator; Chen, Hongda (hongda.chen@usda.gov) – USDA-NIFA.

Brief Summary of Minutes

Members briefly introduced / reacquainted themselves.

Administrative updates (Steve Lommel and Hongda Chen) relayed on-going operational challenges, including severe budgetary cuts at experiment stations in upcoming year, move of USDA-NIFA to Kansas City and ensuing loss of 80% of staff. Nevertheless USDA has invested $16.1 M in competitive research programs relevant to this project, including Foundational and Applied Science priority area A1511 (Nanotechnology for Agricultural and Food Systems) relevant to this project at funding rates of ~$5M in 2017 and 2018. Hongda Chen also promoted the Rapid Response to COVID-19 solicitation ($9 M) with proposals due June 4, 2020.

Administratively projects are asked for more abbreviated (3 page limit) annual reports with stronger emphasis on impacts and products and alignment to societal challenges. Also this project is coming into its final year, so members will need to submit a renewal proposal addressing new directions after reflecting on the team’s expertise, developed collaborations among the members and how these collaborations stimulate new grants that create new avenues for research and training.

Station reports were delivered from each participant in the meeting describing completing and on-going research.

Jose Reyes-de-Corcuera was elected Secretary of the project for the coming year (with Olga Tsyusko advancing to vice-chair, and Daniel Jenkins advancing to chair). The remainder of the meeting was devoted to discussing options for future meetings and coordinating joint projects, publications, and collaborations. Full minutes are available at []

Accomplishments

Over this project year participants made numerous contributions towards achieving the corresponding (year 4) milestones delineated in our original proposal. <ol> <li>Continue to validate the biosensors and other devices in real matrices.</li> </ol> Participants reported numerous advances in nanomaterial-enabled biosensor technologies across a spectrum of applications, and to novel antimicrobial properties of nanomaterials to improve safety and security of the food supply. Notable advances include innovative nanoparticle morphologies for enhanced Surface Enhanced Raman Scattering (SERS) for pesticide analysis (MO), investigating metabolite distributions in plant and animal cells (IA, UT), and to enhance performance of a variety of other biosensing modalities including for microbial and chemical contaminants in food and water (AR, AZ, FL, HI, IA, MI, WI). Participants deepened multidisciplinary collaborations to address antimicrobial resistance, advancing affordable technologies for rapid clinical identification of antimicrobial resistance in pathogenic bacteria (MI), and novel approaches to preventing and treating infections with pathogenic organisms demonstrated successfully in animal models (SC). To support commercialization of new technologies participants contributed to the development of open source smart-phone based approaches for portable analysis (AR, AZ, HI), scalable disposable sensors including using paper microfluidics (AZ), scalably manufactured graphene (FL, IA), as well as stabilization of immobilized biomaterials (GA). At least two groups have made significant progress characterizing and understanding mechanisms of toxicology of a wide array of commonly used nanomaterials (KY, WI). Participants have also engineered new nanomaterials for targeted drug delivery (AR, IA) or vaccines against animal viruses (VA), to make edible films with novel antimicrobial and nutritional properties (NJ), to improve efficiency / rates for value-added bioprocessing (AR, WI), and enhance speed and sensitivity for recovery and detection of pathogenic bacteria (AR, MI). Complementary to work discrete biosensing technologies for wide distribution in the environment, several participants of this project are also working on networking approaches and artificial intelligence to aggregate data and provide meaningful decision support to improve food quality, safety, and the cost-effectiveness of agriculture and bioprocessing (AR, AZ, FL, MI, NJ).   <ol start="2"> <li>Identify potential industry partners and initiate meetings with these partners, and;</li> <li>Assess market-readiness of the technologies.</li> </ol> Several groups partnered in various initiatives to strengthen ties to industry, including identification and execution of new research needs, and to strengthen research capacity globally to develop practical, cost-effective biosensing technologies. These include partnerships with the Global Alliance for Rapid Diagnostics (GARD) to establish centers of excellence (COEs) around the world, and agreements with food companies to validate and license technologies for rapid pathogen extraction / detection (MI). Walmart foundation awarded $3.5 M to a group lead by a project member from Arkansas for research on biosensors for food safety. A consortium of University groups including participants in this project (FL, IA) are actively involved in a funded planning grant under NSF-IUCRC (Industry-University Cooperative Research Center) to use nano-enabled sensory tools to study soil dynamics, and technologies developed by members of our group have successfully been used by government and industry, such as handheld biosensors for bacteria in drinking water used by Tucson Water (AZ). Participants at Florida are working with SBIR funding to develop new mobile device APIs to facilitate risk analysis in water quality. <ol start="4"> <li>Continue the exchange of educational teaching materials on nanotechnology and biosensors among member institutions.</li> </ol> Project members are highly active in instruction, developing and sharing teaching / training materials related to nanotechnology and biosensors. This includes leading two NSF-REU programs (IA) where students contributed to research and development of wearable graphene-based stress sensors. <ol start="5"> <li>Conduct annual meeting to report progress of research activities.</li> </ol> In person reporting of progress from each station was made in an annual meeting virtually on May 21-22, 2020. Minutes of this meeting are at the link above. Outputs of this project this year include at least 76 peer-reviewed publications in high impact journals, and research is regularly highlighted as the feature article in their respective issues, and / or covered in the scientific press (i.e. NSF Research News and CEP Magazine) (AZ). Participants have notably made publicly available several research tools to help identify relevant research (SENSEE; FL) and share protocols for research related to biosensors and antimicrobial resistance (IA, MI). Activities planned for the upcoming year are to collaborate on a review paper on biosensors in food, agriculture, and the environment to help direct the focus of the group, enhance collaborations on interdisciplinary topics related to antimicrobial resistance, and to consider future directions to guide development of a renewal proposal for NC-1194 and to support successful transition of new technologies and ideas to industry.

Publications

<ol> <li>Anu Prathap, M.U., Castro-Pérez, E., Jiménez-Torres, J.A., Setaluri, V., Gunasekaran, S., 2019. A flow-through microfluidic system for the detection of circulating melanoma cells. Biosens. Bioelectron. 142, 111522. https://doi.org/10.1016/j.bios.2019.111522</li> <li>Asgari, S., Sun, L., Lin, J., Weng, Z., Wu, G., Zhang, Y., Lin, M., 2020. Nanofibrillar cellulose/Au@Ag nanoparticle nanocomposite as a SERS substrate for detection of paraquat and thiram in lettuce. Microchim. Acta 187, 390. https://doi.org/10.1007/s00604-020-04358-9</li> <li>Bao, R., Gao, N., Lv, J., Ji, C., Liang, H., Li, S., Yu, C., Wang, Z., Lin, X., 2019. Enhancement of Torularhodin Production in Rhodosporidium toruloides by Agrobacterium tumefaciens-Mediated Transformation and Culture Condition Optimization. J. Agric. Food Chem. 67, 1156–1164. https://doi.org/10.1021/acs.jafc.8b04667</li> <li>Bhusal, N., Shrestha, S., Pote, N., Alocilja, E.C., 2018. Nanoparticle-Based Biosensing of Tuberculosis, an Affordable and Practical Alternative to Current Methods. Biosensors 9. https://doi.org/10.3390/bios9010001</li> <li>Bills, M.V., Loh, A., Sosnowski, K., Nguyen, B.T., Ha, S.Y., Yim, U.H., Yoon, J.-Y., 2020. Handheld UV fluorescence spectrophotometer device for the classification and analysis of petroleum oil samples. Biosens. Bioelectron. 159, 112193. https://doi.org/10.1016/j.bios.2020.112193</li> <li>Bills, M.V., Nguyen, B.T., Yoon, J.-Y., 2019. Simplified White Blood Cell Differential: An Inexpensive, Smartphone- and Paper-Based Blood Cell Count. IEEE Sens. J. 19, 7822–7828. https://doi.org/10.1109/JSEN.2019.2920235</li> <li>Bills, M.V., Yoon, J.-Y., 2020. Label-Free Mie Scattering Identification of Tumor Tissue Using an Angular Photodiode Array. IEEE Sens. Lett. 4, 1–4. https://doi.org/10.1109/LSENS.2020.3001489</li> <li>Briceno, R.K., Sergent, S.R., Benites, S.M., Alocilja, E.C., 2019. Nanoparticle-Based Biosensing Assay for Universally Accessible Low-Cost TB Detection with Comparable Sensitivity as Culture. Diagn. Basel Switz. 9. https://doi.org/10.3390/diagnostics9040222</li> <li>Cai, G., Zheng, L., Liao, M., Li, Y., Wang, M., Liu, N., Lin, J., 2019. A microfluidic immunosensor for visual detection of foodborne bacteria using immunomagnetic separation, enzymatic catalysis and distance indication. Mikrochim. Acta 186, 757. https://doi.org/10.1007/s00604-019-3883-x</li> <li>Cao, X., Zhu, X., He, S., Xu, X., Ye, Y., Gunasekaran, S., 2019. Gold nanoparticle-doped three-dimensional reduced graphene hydrogel modified electrodes for amperometric determination of indole-3-acetic acid and salicylic acid. Nanoscale 11, 10247–10256. https://doi.org/10.1039/C9NR01309A</li> <li>Chen, B., Gsalla, A., Gaur, A., Lui, Y.H., Tang, X., Geder, J., Pruessner, M., Melde, B.J., Medintz, I.L., Shafei, B., Hu, S., Claussen, J.C., 2019a. Porous Wood Monoliths Decorated with Platinum Nano-Urchins as Catalysts for Underwater Micro-Vehicle Propulsion via H2O2 Decomposition. ACS Appl. Nano Mater. 2, 4143–4149. https://doi.org/10.1021/acsanm.9b00593</li> <li>Chen, B., Kruse, M., Xu, B., Tutika, R., Zheng, W., Bartlett, M.D., Wu, Y., Claussen, J.C., 2019b. Flexible thermoelectric generators with inkjet-printed bismuth telluride nanowires and liquid metal contacts. Nanoscale 11, 5222–5230. https://doi.org/10.1039/C8NR09101C</li> <li>Chung, S., Jennings, C.M., Yoon, J.-Y., 2019. Distance versus Capillary Flow Dynamics-Based Detection Methods on a Microfluidic Paper-Based Analytical Device (μPAD). Chem. – Eur. J. 25, 13070–13077. https://doi.org/10.1002/chem.201901514</li> <li>Dachavaram, S.S., Moore, J.P., Bommagani, S., Penthala, N.R., Calahan, J.L., Delaney, S.P., Munson, E.J., Batta‐Mpouma, J., Kim, J.-W., Hestekin, J.A., Crooks, P.A., 2020. A Facile Microwave Assisted TEMPO/NaOCl/Oxone (KHSO5) Mediated Micron Cellulose Oxidation Procedure: Preparation of Two Nano TEMPO-Cellulose Forms. Starch - Stärke 72, 1900213. https://doi.org/10.1002/star.201900213</li> <li>Dieckhaus, L., Park, T.-S., Yoon, J.-Y., 2020. Smartphone based paper microfluidic immunoassay of Salmonella and E. coli, in: Schatten, H. (Ed.), Salmonella: Methods and Protocols. Springer, New York.</li> <li>Dong, Xiufang, Bai, Y., Xu, Z., Shi, Y., Sun, Y., Janaswamy, S., Yu, C., Qi, H., 2019. Phlorotannins from Undaria pinnatifida Sporophyll: Extraction, Antioxidant, and Anti-Inflammatory Activities. Mar. Drugs 17, 434. https://doi.org/10.3390/md17080434</li> <li>Dong, Xiuping, Liu, W., Song, X., Lin, X., Yu, D., Yu, C., Zhu, B., 2019. Characterization of Heat-Induced Water Adsorption of Sea Cucumber Body Wall. J. Food Sci. 84, 92–100. https://doi.org/10.1111/1750-3841.14392</li> <li>Dutta, S.D., Patel, D.K., Seo, Y.-R., Park, C.-W., Lee, S.-H., Kim, J.-W., Kim, J., Seonwoo, H., Lim, K.-T., 2019. In Vitro Biocompatibility of Electrospun Poly(ε-Caprolactone)/Cellulose Nanocrystals-Nanofibers for Tissue Engineering [WWW Document]. J. Nanomater. https://doi.org/10.1155/2019/2061545</li> <li>Eke, J., Mills, P.A., Page, J.R., Wright, G.P., Tsyusko, O.V., Escobar, I.C., 2020. Nanohybrid Membrane Synthesis with Phosphorene Nanoparticles: A Study of the Addition, Stability and Toxicity. Polymers 12, 1555. https://doi.org/10.3390/polym12071555</li> <li>Franco, A.J.DM., Merca, F.E., Rodriguez, M.S., Balidion, J.F., Migo, V.P., Amalin, D.M., Alocilja, E.C., Fernando, L.M., 2019. DNA-based electrochemical nanobiosensor for the detection of Phytophthora palmivora (Butler) Butler, causing black pod rot in cacao (Theobroma cacao L.) pods. Physiol. Mol. Plant Pathol. 107, 14–20. https://doi.org/10.1016/j.pmpp.2019.04.004</li> <li>Gómez-Velasco, A., León-Cortés, J.L., Gordillo-Marroquín, C., Sánchez-Pérez, H.J., Alocilja, E.C., Muñoz-Jiménez, S.G., Bencomo-Alerm, A., Enríquez-Ríos, N., Jonapá-Gómez, L., Gómez-Bustamante, A., 2019. Uso de nanopartículas magnéticas y un biosensor para el diagnóstico y monitoreo de enfermedades infecciosas emergentes, re-emergentes y tropicales desatendidas. Enfermedades Emerg. 18, 23–31.</li> <li>Guo, R., Wang, S., Huang, F., Chen, Q., Li, Y., Liao, M., Lin, J., 2019. Rapid detection of Salmonella Typhimurium using magnetic nanoparticle immunoseparation, nanocluster signal amplification and smartphone image analysis. Sens. Actuators B Chem. 284, 134–139. https://doi.org/10.1016/j.snb.2018.12.110</li> <li>He, K., Li, Z., Wang, L., Fu, Y., Quan, H., Li, Y., Wang, X., Gunasekaran, S., Xu, X., 2019a. A Water-Stable Luminescent Metal–Organic Framework for Rapid and Visible Sensing of Organophosphorus Pesticides. ACS Appl. Mater. Interfaces 11, 26250–26260. https://doi.org/10.1021/acsami.9b06151</li> <li>He, K., Yang, H., Wang, L., Guan, J., Wu, M., He, H., Gunasekaran, S., Wang, X., Wang, Q., Xu, X., 2019b. A universal platform for multiple logic operations based on self-assembled a DNA tripod and graphene oxide. Chem. Eng. J. 368, 877–887. https://doi.org/10.1016/j.cej.2019.03.019</li> <li>Hondred, J.A., Medintz, I.L., Claussen, J., 2019. Enhanced electrochemical biosensor and supercapacitor with 3D porous architectured graphene via salt impregnated inkjet maskless lithography. Nanoscale Horiz. 4, 735–746. https://doi.org/10.1039/C8NH00377G</li> <li>Hu, J., Zhao, T., Li, S., Wang, Z., Wen, C., Wang, H., Yu, C., Ji, C., 2019. Stability, microstructure, and digestibility of whey protein isolate – Tremella fuciformis polysaccharide complexes. Food Hydrocoll. 89, 379–385. https://doi.org/10.1016/j.foodhyd.2018.11.005</li> <li>Jenkins, D.M., Lee, B.E., Jun, S., Reyes-De-Corcuera, J., McLamore, E.S., 2019. ABE-Stat, a Fully Open-Source and Versatile Wireless Potentiostat Project Including Electrochemical Impedance Spectroscopy. J. Electrochem. Soc. 166, B3056. https://doi.org/10.1149/2.0061909jes</li> <li>Kaarj, K., Madias, M., Akarapipad, P., Cho, S., Yoon, J.-Y., 2020a. Paper-based In Vitro Tissue Chip for Delivering Programmed Mechanical Stimuli of Local Compression and Shear Flow. J. Biol. Eng. (accepted).</li> <li>Kaarj, K., Ngo, J., Loera, C., Akarapipad, P., Cho, S., Yoon, J.-Y., 2020b. Simple Paper-based Liver Cell Model for Drug Screening. BioChip J. 14, 218–229. https://doi.org/10.1007/s13206-020-4211-6</li> <li>Kaarj, K., Yoon, J.-Y., 2019. Methods of Delivering Mechanical Stimuli to Organ-on-a-Chip. Micromachines 10. https://doi.org/10.3390/mi10100700</li> <li>Kim, H.-B., Jin, B., Patel, D.K., Kim, J.-W., Kim, J., Seonwoo, H., Lim, K.-T., 2019. Enhanced Osteogenesis of Human Mesenchymal Stem Cells in Presence of Single-Walled Carbon Nanotubes. IEEE Trans. Nanobioscience 18, 463–468. https://doi.org/10.1109/TNB.2019.2914127</li> <li>Kwon, T., Gunasekaran, S., Eom, K., 2019. Atomic force microscopy-based cancer diagnosis by detecting cancer-specific biomolecules and cells. Biochim. Biophys. Acta BBA - Rev. Cancer 1871, 367–378. https://doi.org/10.1016/j.bbcan.2019.03.002</li> <li>Lee, K., Park, H., Baek, S., Han, S., Kim, D., Chung, S., Yoon, J.-Y., Seo, J., 2019. Colorimetric array freshness indicator and digital color processing for monitoring the freshness of packaged chicken breast. Food Packag. Shelf Life 22, 100408. https://doi.org/10.1016/j.fpsl.2019.100408</li> <li>Li, Jiaqi, Cao, L., Li, D., Yu, C., Tan, M., 2019. Carbon dots from roasted mackerel (scomberomorus niphonius) for free radical scavenging. LWT 111, 588–593. https://doi.org/10.1016/j.lwt.2019.05.073</li> <li>Li, Jieran, Rodrigues, S., Tsyusko, O.V., Unrine, J.M., 2019. Comparing plant–insect trophic transfer of Cu from lab-synthesised nano-Cu(OH)2 with a commercial nano-Cu(OH)2 fungicide formulation. Environ. Chem. 16, 411–418. https://doi.org/10.1071/EN19011</li> <li>Li, S., Ma, R., Pan, J., Lin, X., Dong, X., Yu, C., 2019. Combined effects of aging and low temperature, long time heating on pork toughness. Meat Sci. 150, 33–39. https://doi.org/10.1016/j.meatsci.2018.12.001</li> <li>Li, Yuqing, Liu, J., Fu, Y., Xie, Q., Li, Yanbin, 2018. Magnetic-core@dual-functional-shell nanocomposites with peroxidase mimicking properties for use in colorimetric and electrochemical sensing of hydrogen peroxide. Microchim. Acta 186, 20. https://doi.org/10.1007/s00604-018-3116-8</li> <li>Lichtenberg, S.S., Laisney, J., Elhaj Baddar, Z., Tsyusko, O.V., Palli, S.R., Levard, C., Masion, A., Unrine, J.M., 2020a. Comparison of Nanomaterials for Delivery of Double-Stranded RNA in Caenorhabditis elegans. J. Agric. Food Chem. https://doi.org/10.1021/acs.jafc.0c02840</li> <li>Lichtenberg, S.S., Nuti, K., DeRouchey, J., Tsyusko, O.V., Unrine, J.M., 2020b. Efficacy of chitosan/double-stranded RNA polyplex nanoparticles for gene silencing under variable environmental conditions. Environ. Sci. Nano 7, 1582–1592. https://doi.org/10.1039/D0EN00137F</li> <li>Lichtenberg, S.S., Tsyusko, O.V., Palli, S.R., Unrine, J.M., 2019. Uptake and Bioactivity of Chitosan/Double-Stranded RNA Polyplex Nanoparticles in Caenorhabditis elegans. Environ. Sci. Technol. 53, 3832–3840. https://doi.org/10.1021/acs.est.8b06560</li> <li>Liu, X., Wang, Z., Yin, F., Liu, Y., Qin, N., Nakamura, Y., Shahidi, F., Yu, C., Zhou, D., Zhu, B., 2019a. Zinc-Chelating Mechanism of Sea Cucumber (Stichopus japonicus)-Derived Synthetic Peptides. Mar. Drugs 17, 438. https://doi.org/10.3390/md17080438</li> <li>Liu, X., Wang, Z., Zhang, J., Song, L., Li, D., Wu, Z., Zhu, B., Nakamura, Y., Shahidi, F., Yu, C., Zhou, D., 2019b. Isolation and identification of zinc-chelating peptides from sea cucumber (Stichopus japonicus) protein hydrolysate. J. Sci. Food Agric. 99, 6400–6407. https://doi.org/10.1002/jsfa.9919</li> <li>Lu, L., Gunasekaran, S., 2019. Dual-channel ITO-microfluidic electrochemical immunosensor for simultaneous detection of two mycotoxins. Talanta 194, 709–716. https://doi.org/10.1016/j.talanta.2018.10.091</li> <li>Lu, S.-Y., Malekanfard, A., Beladi-Behbahani, S., Zu, W., Kale, A., Tzeng, T.-R., Wang, Y.-N., Xuan, X., 2020. Passive Dielectrophoretic Focusing of Particles and Cells in Ratchet Microchannels. Micromachines 11, 451. https://doi.org/10.3390/mi11050451</li> <li>Lv, J., Yang, Z., Xu, W., Li, S., Liang, H., Ji, C., Yu, C., Zhu, B., Lin, X., 2019. Relationships between bacterial community and metabolites of sour meat at different temperature during the fermentation. Int. J. Food Microbiol. 307, 108286. https://doi.org/10.1016/j.ijfoodmicro.2019.108286</li> <li>Mammadova, N., Summers, C.M., Kokemuller, R.D., He, Q., Ding, S., Baron, T., Yu, C., Valentine, R.J., Sakaguchi, D.S., Kanthasamy, A.G., Greenlee, J.J., Heather West Greenlee, M., 2019. Accelerated accumulation of retinal α-synuclein (pSer129) and tau, neuroinflammation, and autophagic dysregulation in a seeded mouse model of Parkinson’s disease. Neurobiol. Dis. 121, 1–16. https://doi.org/10.1016/j.nbd.2018.09.013</li> <li>McLamore, E.S., Palit Austin Datta, S., Morgan, V., Cavallaro, N., Kiker, G., Jenkins, D.M., Rong, Y., Gomes, C., Claussen, J., Vanegas, D., Alocilja, E.C., 2019. 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Impact Statements

Project supported training of new scientists researching innovative nanomaterials and biosensors (at least 13 PhD and 6 MS students) and enhanced global networking of researchers with work related to nanotechnology and biosensors. Walmart foundation awarded $3.5 M to a group lead by a project member from Arkansas (Yanbin Li) for research on biosensors for food safety. Numerous technologies developed by project participants have been developed that advance key needs to enable broad distribution of biosensors in a wide variety of applications, including speed to detection, ease of use / portability, scalability / manufacturability, sensitivity / selectivity, and data management / analysis. In addition research has resulted in a variety of innovative use of biological and nanoscale materials for vaccine development, drug delivery and therapies against pathogenic organisms. Toxicological data on widely used nanomaterials can inform policy decisions related to manufacturing and exposure to these materials to safeguard public and environmental health.

Date of Annual Report: 03/27/2022

Report Information

Annual Meeting Dates: 08/09/2021 - 08/10/2021
Period the Report Covers: 10/01/2020 - 09/30/2021