University Of Mississippi

NIH Research Projects · FY 2025 · 2025-01

Project Summary The Mississippi ESTEEMED Scholars Program is designed to promote equitable outcomes for undergraduates in bioengineering/STEM fields who come from underrepresented backgrounds. Mississippi has the highest poverty rate in the nation and grapples with educational challenges, including having most of the state designated as a critical needs area for mathematics and science education. Thus, the pathway to a biomedical engineering career is challenging. To help address these issues, we propose the ESTEEMED program to facilitate the success of incoming freshmen students pursuing biomedical engineering (BME). The program focuses on supporting BME curriculum preparation and building scientific identity, ensuring that students, particularly those from underrepresented backgrounds, are well-equipped to navigate the demanding curriculum and persist through to graduate STEM careers The long-term goal is to train the next generation of scientists that will transform understanding of disease prevention, detection, diagnosis, and treatment through engineering and diversify the biomedical workforce. Transitional programming designed to increase recruitment, retention, and research success of young scholars affiliated with the University of Mississippi will increase the preparedness of future doctoral degree applicants in fields like bioengineering and bioimaging. The program includes a summer bridge that transitions students from high school into the University of Mississippi. They will get a head start on coursework, be trained in responsible conduct of research and laboratory safety, be introduced to foundations of electronics, conduct 3D printing, and be exposed to scientific equipment across numerous facilities. During the academic year, mentorship meetings with program leaders will occur, and individual development plans will be utilized to plot a course towards a career trajectory in STEM for the students. Supplemental self-paced computer programming training will be provided to enhance the preparedness for future coursework, projects, and careers. In addition, students will choose from numerous local research mentors who run independent laboratories that span fields in biomedical engineering, bioimaging, biology, chemistry, physics, and pharmacology. In their second summer, students will receive support to apply to work in external laboratories with highly specialized skills. In addition, during the academic year, students will have the opportunity to interact directly with successful scientists and engineers in multiple engaging formats. Training will be provided to support student presentations of their own laboratory projects to visiting scholars and at local symposia/regional conferences. Finally, this program will support students to transition into an honors program on campus for their junior/senior years to continue their trajectory toward graduate school in STEM fields.

NSF Awards · FY 2024 · 2024-12

Many natural and engineering flows, like wind gusts, water surges, or air moving over airplanes during maneuvers, can rapidly change speed and direction, i.e., under pulsation. These changes can be difficult to predict for flows over rough surfaces as surface protrusions create chaotic swirling and fluctuating forces in response to the shifting flow. Current prediction tools often assume smooth surfaces and steady conditions, leading to prediction errors or designs that work well in ideal scenarios but fail in real-world situations. The proposed project aims to deepen the understanding by conducting high-accuracy simulations that reveal detailed flow physics in pulsating flow over rough surfaces and develop new tools with unprecedented real-time prediction accuracy and off-design adaptability. The research will be integrated with comprehensive engagement initiatives that involve undergraduate students, community college students, and high schoolers to spark interest and cultivate skills in STEM. Outreach activities at a museum and Grade 5-12 schools will deliver accessible fluid-related STEM resources and captivating content to engage and educate the broader public. The proposed project aims to tackle the knowledge and tool gaps via three perspectives: establishing a comprehensive high-resolution dataset of pulsating rough-wall flows, discovering new physical insights via novel statistical analysis, and conducting physics-based predictive modeling. The goals are to: 1) advance the understanding of pulsatile flows over roughened surfaces and how we characterize temporal and spatial coherence of turbulent boundary layers in general; 2) develop a novel set of accurate tools to quantitatively analyze and predict roughness effects in pulsatile flows. Direct numerical simulations and large-eddy simulations will resolve the roughness-scale flow to elucidate previously unexplored flow physics with direct measurements of wall stresses and velocity within the roughness region. Three roughness categories of increasing complexity, ranging from homogeneous to multiscale heterogeneous, will be addressed. In parallel, analytical physics-based models will be developed to explicitly account for drag production by roughness. Simulations will be complemented by collaborative experiments and industrial partnerships to establish a comprehensive research chain facilitating the practical implementation of findings. The improved predictive capability will reduce the design, operation, and maintenance costs of engineered products across aviation, energy, mechanical, meteorological, and environmental industries. Education activities will broaden the participation of individuals in STEM education and research, which will promote education and career readiness, expand the talent pool, and contribute to economic development. This project is jointly funded by Fluid Dynamics program and the Established Program to Stimulate Competitive Research (EPSCoR). This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-10

The objective of this Civic Innovation Challenge Planning Grant (CIVIC-PG) is to support research on multi-hazard risk analysis, which involves the examination of multiple hazards in a specific geographic area and time, their magnitude, their interactions, and the interpretation of their combined effects on the local communities. Many parts of the United States have seen an increasing number of weather and climate disasters, from severe storms and floods to heat waves and droughts. The Mississippi Delta is particularly vulnerable to such disasters, which significantly impact the natural environment and anthropogenic resources. Vulnerability is driven by a combination of factors, including economically disadvantaged and marginalized populations, systemic issues, limited resources, and lack of understanding of risk. This project envisions building disaster-resilient communities by developing advanced, community-driven, open-source multi-hazard risk assessment tools. These tools are critical to developing effective strategies for disaster risk reduction, infrastructure design, urban planning, climate change adaptation, and sustainable economic development. The project establishes a robust partnership between the university and the community to identify social, economic, political, and governance factors that hinder the resilience of the Mississippi Delta communities. Researchers and stakeholders co-develop methods to incorporate local knowledge and contextual understanding of multi-hazard risk into research, policy, and practice. Using web interactive mapping applications, the project harnesses the potential and capabilities of remote sensing and open geospatial data, machine learning algorithms, and cloud computing for risk identification and visualization. The project also addresses the effect of spatial scale on the quantification of multi-hazard risk and researches a fractal-based modeling framework for multi-scale risk assessment. This project is in response to the Civic Innovation Challenge program’s Track A. Climate and Environmental Instability - Building Resilient Communities through Co-Design, Adaption, and Mitigation and is a collaboration between NSF, the Department of Homeland Security, and the Department of Energy. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-10

This project seeks to protect the integrity of digital content and maintain public trust. The rapid advancement of generative Artificial Intelligence (AI) has made it easier to create and manipulate digital content. Current tools for detecting AI-generated content are fragmented and challenging to use. The project team is developing an all-in-one digital content forensics platform designed to streamline the forensic analysis process. By integrating multiple tools into a single platform, it aims to empower users by providing a reliable and user-friendly platform for evaluating digital content. The project employs a user-centered design process, involving extensive qualitative interviews and user studies to understand needs and workflows. Based on the findings of these studies, the team is integrating various digital content forensic tools into a single platform, supported by a robust organization for the coherent navigation and selection of the tools. The team is also exploring explanation methods to enhance user comprehension of each tool’s outputs. Finally, to help users make the most of this platform, the team is creating novel game-based training scenarios and comprehensive ethical frameworks based on professional norms. The team is disseminating this work and other information about the project through workshops and professional networks in multiple user communities that will be able to leverage this platform to maintain the integrity of online digital content. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2024 · 2024-09

Project Summary The 70-kilodalton heat shock protein (HSP70) family, universally conserved across prokaryotic and eukaryotic organisms, serves as essential molecular chaperones for maintaining protein homeostasis. They are central to the pathology of various diseases, including cancer, infectious diseases, neurodegeneration, and cardiovascular disorders. Beyond their established intracellular roles, HSP70 proteins also manifest in the extracellular space, significantly influencing cancer biology by modulating immune responses and facilitating metastasis. This positions them as vital biomarkers for deciphering and potentially targeting the mechanisms underlying cancer pathology. The critical involvement of HSP70 in such diverse human pathologies underscores the potential of therapeutic strategies to modulate HSP70 pathways. Delving into the complex structure-function relationship of HSP70 is essential for developing novel therapeutic interventions capable of addressing various pathologies. This project aims to elucidate the intricate roles of the cytosolic HSP70, with a particular emphasis on its underexplored structural attributes and their ramifications for cellular operations and disease pathogenesis. The long-term objectives include a deeper understanding of HSP70’s role in maintaining cellular integrity and stress response, which is crucial for developing innovative therapeutic interventions for various diseases. Specifically, the research will concentrate on HSP70’s lectin-like capacity to bind O-GlcNAcylated proteins, its ability to bind calcium/calmodulin, and its extracellular functions. These functionalities are hypothesized to be pivotal in modulating cellular thermoresistance and stress responses and potentially influencing the onset and progression of various diseases. The chosen model system for this investigation is Arabidopsis cytosolic HSP70 (AtHSP70), selected for its significant homology to human HSP70 isoforms and its established role in conferring heat stress resilience in plants. This model provides a robust framework for dissecting the conserved mechanisms of HSP70 function relevant to human health, thus directly aligning with the mission of improving the understanding and treatment of human diseases. To achieve these aims, the project will employ cutting-edge techniques such as proximity labeling with TurboID to identify novel protein interactions in vivo and lectin weak affinity chromatography to characterize the interaction between HSP70 and glycosylated proteins. Through a detailed examination of these interactions, the project seeks to expand the current understanding of HSP70’s intracellular and extracellular functions and uncover potential therapeutic targets within the HSP70 pathway. By revealing how alterations in HSP70’s interactions with carbohydrates and Ca2+/CaM can influence disease mechanisms, the research aims to open new avenues for developing therapies that modulate these pathways. In essence, this project embodies a strategic approach to bridging fundamental biological research with potential clinical applications, thereby underscoring its relevance to the overarching goal of advancing human health by targeting the molecular bases of disease.

NIH Research Projects · FY 2025 · 2024-09

By third grade, poor science achievement among youth may pave their path to poor adult health, fewer employment opportunities, and lower economic mobility. LEARN Science – Library Efforts to Advance Reading and Nutrition Science – blazes a new trail for youth using informal science education (ISE) through community public libraries across Mississippi (MS). LEARN Science is a life science and reading program through a nutrition science lens to improve science achievement among children in kindergarten through 2nd grade (K-2). MS boasts a robust public library system. Public libraries are potential: 1) bridges to improve Science, Technology, Engineering, and Mathematics (STEM) opportunities; and 2) key collaborators for health education and promotion. The role of food for survival is a foundational scientific concept for children to learn in K-2. Food exposes youth to potentially dozens of scientific concepts daily, as well as to cultural traditions and health principles. As such, nutrition education can empower youth to make food choices for chronic disease prevention. Our overall long-term goal is to establish LEARN Science to improve science achievement among children in K-2. The program will be aligned with Next Generation Science Standards (NGSS) and be consistent with the social ecological model. LEARN Science will consist of seven lesson units. Each unit will follow a storyline and blend two age-appropriate books with: 1) nutrition science terminology; 2) inquiry-based, hands-on activities using science tools; 3) scientific observations and reflection; and 4) take-home family learning resources. Unit readings will include both: 1) a narrative (fiction) book with a nutrition component; and 2) an informational (nonfiction) book showcasing a scientist and/or a scientific principle. Units will include a mentor guide and information on connecting families with community food resources. We will: 1) develop and evaluate the LEARN Science program, utilizing latest educational best practices; 2) establish the NIH SEPA Library Academy, a professional development program (PDP) focused on mentoring and building self-efficacy among library staff to deliver LEARN Science across MS; and 3) disseminate LEARN Science across the MS public libraries. LEARN Science will strengthen ISE opportunities for children in K-2; improve science achievement of children; and ultimately has the potential to expand the number of children choosing STEM careers.

NIH Research Projects · FY 2025 · 2024-09

The proposed training program aims to address the lack of specialized training in applying data science methods, including artificial intelligence (AI)/machine learning (ML), for addiction research at Mississippi’s major universities. The program is featured by its evidence-based, multimodal approach and extensive interdisciplinary collaboration. The design of a progressive learning pathway facilitates learning access for students from various backgrounds. A series of webinars will help raise campus-wide awareness and interest in the topic, laying the groundwork for the recruitment and participation of undergraduate and graduate students in a summer academy that provides hands-on training and subsequent mentored research experiences using data science methods for addiction research. Students participating in the summer academies and mentored research experiences will be encouraged to develop research projects suitable for academic conferences and/or journal submissions. The program will also establish a dedicated research lab to serve as a central hub for collaborative mentoring, program event announcements, and learning resources distribution. Furthermore, an e-learning platform will be used to broaden the program’s reach, and faculty members will work to integrate training content into course curricula to enhance both its impact and long-term sustainability. Successful implementation of the program will equip students to apply data science methods for addiction research, inspire pursuit of advanced degrees in the field, foster cross-disciplinary and institutional collaboration, and strengthen the field’s capacity to address complex addiction challenges.

NSF Awards · FY 2024 · 2024-09

The practice of charging different prices to different consumers, i.e., price discrimination, is commonly used by firms in a variety of industries with the intention of increasing profits. Yet, little is known about the impact on consumers of modern sophisticated discriminatory pricing practices that leverage increasingly detailed information. This gap in understanding must be overcome to develop rules that balance positive aspects of these practices like efficiency against redistributive harm and privacy concerns. The researchers study these issues in the context of two different strategies used in the airline industry: personalized pricing via targeted discounts and auctions for upgraded services and amenities. The empirical analysis relies on unique data obtained from a North American airline to gain deeper insights into the practice of price discrimination by evaluating current and potential price-discrimination strategies. The results of the analysis provide insight into the effectiveness of different strategies and methods on profit and consumer welfare, while also offering guidance to decision makers on how to limit any negative impacts on consumers. The research contributes to knowledge of the welfare impacts of modern discriminatory pricing practices, as well as the challenges associated with implementation. The detailed proprietary data used in the empirical analysis provides a unique opportunity to study two different practices used in the airline industry: personalized pricing via targeted discounts and upgrade auctions. The results of the analysis regarding firms’ ability to leverage increasingly detailed data and implement sophisticated mechanisms to price discriminate is directly of interest to economists and important for development of rules to limit harm to consumers. The contribution of the research is in the field of econometrics and industrial organization, and specifically advances econometric methods for personalized pricing and integration of auctions into a dynamic-pricing environment. These advancements provide a framework for studying these topics in other industries, and guidance for important discussions around data privacy and price discrimination. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

The University of Texas at Austin and the University of Mississippi are conducting a research synthesis study of the barriers and solutions in postsecondary STEM training settings for students with disabilities. The lack of full inclusion of people with disabilities in the STEM workforce is a missed opportunity to realize the full potential and talent of the entire U.S. population. Opportunities to advance knowledge about STEM postsecondary training setting barriers and solutions for students with disabilities will lead to increasing the engagement, academic retention, degree completion, and career advancement of undergraduate students with disabilities in STEM. Such success is essential for building and advancing a robust U.S. STEM workforce. The research team is engaging with an expert advisory board, and postsecondary students with disabilities, to contribute to the synthesis project work. The systematic review includes analyses that are informed by theoretical frameworks and conceptual models that have been used to study the experiences of STEM undergraduate students with disabilities. Published findings include the rationales for decisions made throughout the systematic review and analysis so that others may replicate the synthesis research. This award has been made in response to the NSF solicitation “Workplace Equity for Persons with Disabilities in STEM and STEM Education” (NSF 23-593). The project is funded by the Division of Equity for Excellence in STEM’s Hispanic Serving Institutions program (HSI), the Division of Equity for Excellence in STEM’s Louis Stokes Alliances for Minority Participation program (LSAMP), and the Established Program to Stimulate Competitive Research (EPSCoR). This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

Advances in artificial intelligence (AI) have the potential to rapidly transform jobs, organizations, leisure, social life, health care, education, industry, domestic politics, and international relations. American colleges and universities are developing a variety of courses and modules to ensure that students gain not only the technical competencies needed to develop, understand, deploy, and use AI but also the ethical competencies needed to ensure that these advances are used wisely to contribute to a more productive workforce and a stronger, fairer, and more prosperous nation. Despite the rapid expansion of AI ethics education interventions across various institutions, there is a notable absence of empirical research systematically mapping or comparing these interventions. To address this gap, this project aims to conduct the first-of-its-kind national survey on the state of AI ethics education interventions and how faculty and administrators, as well as their institutions, approach AI ethics education. A key aspect of the research is the development of meaningful collaborations between the three R1 universities and regional institutional partners with diverse stakeholders. The research will be conducted through three regional networks, each anchored by an R1 institution that connects area higher education institutions (HEIs) such as (minority-serving institutions (MSIs), community colleges, and research-intensive universities) and actively engages them in the design, implementation, and dissemination of research. Using a variety of methods (e.g., quantitative surveys and qualitative interviews with faculty and administrators, as well as natural language processing analysis of survey and interview data), the project team will analyze the state of AI ethics interventions in diverse institutions across the United States by (a) mining existing interventions to produce a comprehensive overview of current and planned AI ethics education; (b) developing a framework for describing the ways in which the faculty perceive and conceptualize AI ethics education; (c) exploring the factors that affect the decision- making of instructors while proposing, designing, and offering various AI ethics-related interventions; and (d) identifying institutional capacity and needs to support effective AI ethics education. Overall, the research will allow STEM faculty and educational researchers to craft curricula and administrators to develop institutional initiatives that generate AI ethics competencies tailored to the needs of their students, their employers, and their communities. This project is jointly funded by the Directorate for Engineering, the Directorate for STEM Education, and the Directorate for Computer and Information Science and Engineering, and is managed by the Division of Engineering Education & Centers on behalf of the ER2 Program of the Directorate for Social, Behavioral and Economic Sciences. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-09

Biological evolution is possible only when variation exists within a species. In order to better understand such biological change, ideally both the genetic basis of these variations and the evolutionary forces acting on them need to be well characterized. However, until recently, such research has been impossible or prohibitively costly. Leveraging a fascinatingly variable species and recent technological advances, the goal of this research project is to better understand the evolutionary drivers of diverse phenotypes (appearances) among individuals within a single species. In order to explore this phenomenon of phenotypic diversity the project will focus on a species of poison frog for which phenotype is of critical importance to the survival of the individual by warning away predators. This project will identify the genetic factors responsible for phenotypic differences among individuals so that both the means by which genetic mutations may effect these changes and the forces of evolution that permit such variation can be better understood. The project will include the training of a diverse group of undergraduate students and outreach to area primary and secondary schools. In order to fully appreciate the role of population level phenomena (natural and sexual selection) on the process of interpopulation diversification, this work seeks to characterize the demographic structure of populations of the poison frog species Dendrobates tinctorius. This species is of particular interest because of the extreme degree of interpopulation variation observed in its color and pattern, traits that have been shown to have evolved to warn predators of the frog's toxicity. As an aposematic species, the radical phenotypic polytypism observed in this species seems counterintuitive. This research will leverage a newly-available, high-quality genome assembly for D. tinctorius with new genome sequences of >100 individuals from throughout the geographic range to better understand the population structure of this polytypic species. Further, these new low- and medium-coverage genomes will be used to explore the genetic basis of phenotypic traits associated with the aposematic signal of D. tinctorius (color, pattern). Together these data will provide unprecedented insight the interplay between selection and population structure in the process of evolution and speciation. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

This collaborative project between seven institutions establishes the Mississippi Nano-bio and ImmunoEngineering Consortium (NIEC) to enhance biomaterials research, education, and workforce development in Mississippi. Partnering institutions include Alcorn State University, Jackson State University, Mississippi State University, Tougaloo College, the University of Mississippi, the University of Mississippi Medical Center, and the University of Southern Mississippi. NIEC will develop new materials, test their interactions with biological systems, and evaluate their effectiveness in treating diseases, with a focus on addressing health disparities in Mississippi and the region, ensuring long-term benefits to the state, region, and country. The project's goals include synthesizing and characterizing novel biomaterials appropriate for safe clinical use, educating and retaining a diverse group of scientists and engineers in Mississippi, and securing sustained funding to advance this work. By building a comprehensive research network, promoting inclusivity, and supporting local biotech startups, NIEC seeks to impact science, healthcare, and the state's economy, creating high-tech, well-paying jobs in Mississippi and fostering economic growth. The project aims to create a robust pipeline of next-generation materials by fostering a collaborative, multidisciplinary research team centered around three research focus areas (RFAs): (i) developing biomimetic materials to modulate nano-immuno interactions via protein corona engineering, (ii) designing polymer nanocarriers for efficient nucleic acid complexation and release, and (iii) developing pathogen resilient bioinspired polymeric scaffolds for tissue regeneration. Leveraging NIEC's expertise in nanomaterials synthesis, physicochemical characterization, and computational modeling, these RFAs will form an integrated design loop that will enhance understanding of the interface between biological systems and nanomaterials, establishing generalizable structure-property-function relationships that support the safe and effective translation of innovative biomaterials into clinical applications. In addition to the three biomaterials RFAs, an evaluation of state policies and regulations influencing the growth of the biotech industry in Mississippi will be conducted. This project will measurably impact the preparation of a diverse research-ready workforce that can foster the economic development in MS through (i) leveraging existing undergraduate, graduate, postdoctoral, and earlier career faculty training; (ii) strategically hiring faculty, providing seed grants, and offering mentoring for junior faculty and postdoctoral researchers; and (iii) promoting the development of intellectual property and its commercialization. This project is funded by the NSF EPSCoR Research Incubators for STEM Excellence (E-RISE) Research Infrastructure Improvement Program. The E-RISE RII program supports the development and implementation of sustainable broad networks of individuals, institutions, and organizations that will transform the science, technology, engineering and mathematics (STEM) research capacity and competitiveness in a jurisdiction within a field of research aligned with the jurisdiction's science and technology priorities. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Natural products produced by bacteria are a critical resource for discovery of new therapeutics. Synthetic biology approaches such as heterologous expression of biosynthetic gene clusters (BGCs) have leveraged growing capabilities to assume molecular details of natural products produced by sequenced bacteria from genetic data. Methodologies for heterologous expression of BGCs from Streptomyces spp. have benefited from incredible progress and enabled continued discovery of biologically active metabolites from Gram-positive actinobacteria. However, comparatively fewer synthetic biology techniques are available to other groups of bacteria known to be reliable sources of therapeutic leads. Myxobacteria are considered the Gram-negative reciprocal to actinobacteria when describing gifted producers of natural products that have large genomes replete with BGCs. More importantly, metabolites discovered from myxobacteria occupy chemical space unique from actinobacterial metabolites. The long-term objection of this proposal is to increase the synthetic biology toolset available for natural products discovery from myxobacteria. Expanding the number of heterologous host chassis beyond the lone host Myxococcus xanthus and developing novel strategies to generate host platforms will afford alternative approaches to utilize BGCs for natural products discovery. We aim to: 1) assess alternative myxobacterial plasmids and heterologous hosts, 2) develop a predator-prey model system to observe horizontal transfer of BGCs and explore predation-based methods to engineer heterologous hosts, and 3) apply developed myxobacterial tools to enable discovery of novel bacterial metabolites. Successful completion of proposed research will expand synthetic biology tools suited for discovery of natural products and increase investigation of myxobacterial metabolites with antibacterial, antifungal, cytotoxic, antiviral, and immunosuppressive activities. Sustained access to natural products from bacteria benefits human health by providing therapeutic leads, increasing biosynthetic discoveries, and enabling subsequent clinical pharmacology.

NSF Awards · FY 2024 · 2024-07

Lignite is a low rank of coal that is abundant in most Gulf Coastal plain aquifers and has been shown to cause increased occurrences of kidney disease on the Balkan Peninsula of eastern Europe and in Texas, Arkansas, and Louisiana. Mississippi led the nation in kidney disease mortality from 2015-2020 and was second in the nation in 2021. This early concept research project will collect water chemistry data throughout the state of Mississippi and will perform a suite of geospatial statistical analyses on the collected well water data and existing datasets from the US Geological Survey and from the medical sector and other sources to see if ground water from Gulf Coastal Plain aquifers can be linked to kidney disease in Mississippi. Broader impacts of this research is relevant for all residents of Mississippi and potentially residents of all Gulf Coastal Plain states who obtain their water from ground water wells. If a correlation is found, the results of this research can be used to develop mitigation strategies and could identify zones within specific aquifers or specific geographic regions that should be avoided as domestic water supply sources. Climate-related drought and unpredictable rainfall patterns have led to increasing reliance on groundwater for agricultural irrigation in the Gulf Coastal Plain resulting in significant increases in annual withdrawals from aquifers that are also used for domestic water supplies via municipal and private drinking water wells. Most Gulf Coastal Plain aquifers have naturally occurring lignite, which has been shown to cause increased rates of kidney disease in other parts of the US and world. Mississippi’s mortality rates for kidney disease are among the highest in the nation; but it is unknown if this high rate may be related to the presence of lignite in Gulf Coastal Plain aquifers or due to some other cause. This project applies a variety of machine learning algorithms and techniques on datasets that include water well construction data (location, depth of screened interval, age), health data from the State of Mississippi Medicare database, U.S. Census data, and geochemistry results from a new reconnaissance groundwater survey this project will complete. Goals are to see if there is a statistical correlation between increased rates of kidney disease and the use of lignite-bearing ground water aquifers. Kidney health data will be filtered and classified by a medical doctor specializing in renal functions. Analyses will initially be done for each county in the state. The project funds a Masters-level student in geology at the University of Mississippi which is in an EPSCoR state. The student will learn multidisciplinary skills in geoscience and human health, including: (1) importing and managing data in a geographic information system, (2) performing geospatial analyses techniques; (3) learning how to integrate datasets into machine learning algorithms and how to train AI for neural network analyses on a supercomputer; (4) engaging and interacting with a large cross section of society through seeking permission to sample domestic wells throughout the state; and (5) sampling protocols and techniques for collection of data from water wells, including maintaining chain of custody documentation. Project results may have implications for millions of residents in states across the Mississippi Gulf Coastal Plain states (i.e., Alabama and Georgia) who use groundwater for drinking water. It many also have impact on states such as West Virginia, Kentucky, and Pennsylvania with significant coal-mining industries that have aquifers where there is an intersection of domestic water sources and coal mining activities. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2024-07

Abstract The Zhu lab uses synthetic chemistry to answer questions related to post-translational modifications (PTMs) of proteins, specifically, O-glycosylation and Tyrosine sulfation. Co-localization of Tyrosine sulfation and O- glycosylation on Serine/Threonine (CSOG) is an emerging global PTM pattern. CSOG exists in a variety of proteins, like G-protein coupled receptors, proteins of the hemostatic system, pathogen encoded proteins, and adhesion molecules. CSOG plays critical roles in modulating the binding/signaling networks through those proteins and their binding ligands/proteins. However, detailed information about how the cells or biological systems use fine-tuning CSOG structures to modulate corresponding binding/signaling pathways is lacking. One of the main gaps for studying the relationship is the lack of structure-defined molecules bearing CSOG. To fill up the gap, a generalized strategy is needed to synthesize those molecules so we can get access to the massive, diversified structures of CSOG. Then we can map out the relationship between the fine-tuning structures of CSOG and their corresponding binding/signaling pathways. We can also use these structure-defined molecules to develop reliable methods for characterizing CSOG from different proteins. Furthermore, we can combine synthetic chemistry and computational chemistry to develop robust computational chemistry methods for predicting meaningful bindings and designing probes/molecules to target certain specific bindings. Our lab currently focuses on applying site-selective sulfation in chemoenzymatic synthesis of glycopeptides to provide a generalized strategy for the access to the complex molecules with CSOG. Towards this goal, our lab developed a Ag2O promoted method for large-scale synthesis of the two critical intermediates, GalN3-α-Fmoc- Ser/Thr, for future synthesis of all the needed 16 glyco-amino acids. We have achieved the synthesis of core 2 containing glyco-amino acid in gram scale. For site-selective sulfation, we developed 2-Cl-trityl protecting group as a new way for doing site-selective sulfation. For enzymatic extension of O-glycans on sulfoglycopeptides, our initial attempt demonstrated that the enzyme worked on the sulfoglycopeptide even with protecting groups on the sulfate groups. We expect to generalize this strategy to synthesize libraries of sulfoglycopeptides in this proposal. This MIRA funding will support us to use our generalized synthetic strategy to initiate the following major projects: (1) Build compound libraries to map out the complex network between COSG patterns and corresponding binding/signaling pathways; (2) Use structure-defined molecules with CSOG to develop reliable methods to characterize CSOG structures from different proteins; (3) Use our binding data and computational methods to develop robust computational chemistry methods to predict meaningful binding and design probes/molecules to target specific bindings.

NIH Research Projects · FY 2025 · 2024-07

ABSTRACT This proposal will address the critical need to define for cannabinoid exposure: 1) the most sensitive developmental exposure windows; 2) the dose- and sex-dependence of adverse outcomes; 3) developmentally relevant molecular mechanisms of persistent adverse effects; and 4) the relative developmental toxicity of other cannabinoids available to consumers. Cannabis/∆9-tetrahydro- cannabinol (THC) is the most commonly used illicit drug by pregnant women, and cannabidiol (CBD) is readily available over the counter with suggested benefits in pregnancy for morning sickness, stress, and sleeplessness. Similarly, other minor cannabinoids are marketed directly to consumers with numerous health claims. Because of maternal use, pre- and post-natal cannabinoid exposures occur during critical stages of children’s brain development despite our lack of understanding of the acute and long-term consequences. In addition, cannabinoid exposure (e.g. via vaping) frequently occurs during adolescence, another sensitive time of brain development and neuronal pruning. In our zebrafish model system, we have observed persistently altered adult behavior after embryos were exposed to THC and CBD during early development. Our central hypothesis is that exposure to cannabinoids causes alterations in inflammatory mediators in the developing brain leading to the persistent alterations in behavior throughout development and into adulthood. Our research framework, depicted as an adverse outcome pathway (AOP), will specifically investigate three aims to measure: 1) morphological and persistent behavioral alterations in anxiety/locomotion; 2) time-of-exposure susceptibilities; and 3) neuroinflammation resulting from THC and CBD developmental exposure. We will use the highly relevant, predictive, and high throughput zebrafish model, including three transgenic lines, to assess the spatial and temporal relationships between microglial response, gene expression, and persistent behavioral adverse outcomes. Cannabinoid-mediated changes in neuroinflammation gene/protein expression and altered cellular trajectories will be identified using single nucleiRNAseq and LC-MS/MS protein validation in larval and adult brains. The dependence of the specific cannabinoid receptors in observed toxicities will be determined by using cannabinoid receptor 1 and 2 null lines. Our ongoing and productive collaborations, including work with three NIH-COBRE core facilities, will be leveraged to inform the developmental origins of health and disease caused by cannabinoid exposure. The proposed research is significant because it will provide new, relevant information to guide cannabinoid policy and healthcare decisions made by pediatricians, obstetricians, and policy- makers needed to ensure public health and safety.

NSF Awards · FY 2024 · 2024-07

Sensor networks play pivotal roles across civilian and military domains, from facilitating cognitive radios in 6G wireless communications to environmental monitoring and military surveillance. These networks often rely on battery-operated sensors that wirelessly transmit data to a fusion center for analysis, enabling the detection of critical phenomena through distributed detection methods. The effectiveness of these systems hinges on achieving high detection performance over noisy wireless channels, while conserving energy. Traditional approaches transmit either raw observations or their summaries to the fusion center. However, they either require higher transmission bandwidth or are not optimized for energy conservation. This project aims to develop and optimize the transformation of raw sensor observations for optimal detection performance over noisy wireless channels, subject to constraints on transmission power and bandwidth. Anticipated outcomes include near-optimal distributed detection solutions, especially in the context of 6G and wireless sensor network deployments, elevating detection precision and facilitating timely decision-making. Additionally, the project seeks to enhance STEM education by providing research opportunities to underrepresented groups, including domestic minority and women students in Mississippi. By exploring alternative data forms beyond the current options of transmitting either original observations or binary codewords, this project aims to determine the most effective means of conveying information from the sensors to the fusion center. This objective will be pursued through a comprehensive investigation into the transformed distributed detection paradigm with several concerted efforts. First, the project focuses on the transformed, quantized distributed detection, assessing its design tractability and identifying its optimal operational region compared to conventional methods under the same bandwidth and power constraints. Second, employing functional analysis and numerical techniques, the project aims to optimize the sensor observation transforms to maximize the distributional divergence for signals received at the fusion center under different hypotheses. Third, the project evaluates the performance of transformed distributed detection in scenarios involving both channel noise and correlated observations, considering both Gaussian and non-Gaussian copula models. By addressing some fundamental issues in the field of distributed detection, including the optimality of the transforms and performance boundaries, the project is expected to yield optimized energy-efficient transmission of sensor information for decision making in networked systems. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2022-08

Project Summary: Actin filaments and microtubules are cytoskeletal polymers essential for cell division, motility, and intracellular transport, and deficiencies in these proteins are implicated in cancer, heart disease, and other disorders. In order to facilitate vital tasks that span the entire cell, these filaments coordinate with each other through motor proteins, such as kinesin and myosin, and associated binding proteins. The molecular basis for this communication through tension and compression forces and how these signals propagate through the cytoskeleton is not well understood. Approaches to study such cytoskeletal phenomena have traditionally been either at the single molecule level or whole cell level, and the actin and microtubule cytoskeletons have generally been evaluated as separate systems in vitro. While single molecule experiments, with methods such as optical trapping, have been invaluable in deciphering the mechanics of individual motors, a completely reductionist approach with one filament and one motor protein does not accurately represent the structural hierarchy in which crosslinking motors and proteins function. On the other hand, cell level studies take place in a quite complex environment. In this research plan, we will bridge the gap in scale and assay control by engineering novel, physiologically relevant cytoskeletal environments, or nanocells, in which to probe motor protein mechanics and cytoskeletal crosstalk. Much like LEGOs, we can choose which cytoskeletal elements to incorporate in our nanocell’s architecture and tune the building blocks accordingly to understand how changes at the molecular level propagate to system level force generation and network stiffness. Using this innovative approach, our overarching goal is to provide a fundamental molecular understanding of how motors, crosslinkers, filaments, and signaling factors communicate with each other in ensembles and to the local cytoskeletal environment utilizing optical trapping, quartz crystal microbalance with dissipation, and spectroscopic techniques. Specifically, we will investigate how myosins work together in ensembles in actin assemblies and what molecular components dictate productive force generation. Hybrid nanocells that consist of elements from both the actin and microtubule cytoskeleton will be probed to understand how polymers of different stiffnesses, crosslinking proteins with different pliability, and motor proteins with varying processivity and force generation capability affect cytoskeletal crosstalk. As E-hooks are the diversity site of tubulin and uniquely influence motility in disparate kinesin families, we will interrogate how E- hook structure affects ensemble kinesin force generation in nanocells. The proposed research will pave the way to our long-term goal, which is not only to understand fundamental mechanisms that sustain life, but ultimately be able to reconstitute physiologically realistic models of cellular processes in vitro, providing an enormous potential for developing diagnostic and treatment strategies for cytoskeletal diseases.

NIH Research Projects · FY 2024 · 2022-03

PROJECT SUMMARY This five-year plan for a MOSAIC K99/R00 Postdoctoral Career Transition Award will equip Dr. Erica O’Brien with essential knowledge and skills to help launch her career as an independent scientist in health- focused aging research. This qualified candidate seeks to focus her research program on understanding the psycho-social and social cognitive determinants, multi-factorial pathways, and temporal dynamics underlying lifelong engagement in health-promoting behaviors and activities. The career development plan involves scientific training at Pennsylvania State University’s College of Health and Human Development under the mentorship of world-renowned experts in ecological momentary assessments, physical activity, aging/adult development, and health behavior research. The proposed training will supplement Dr. O’Brien’s existing background in studying age-related differences in cognitive engagement and in using diary designs with a foundation in intensive longitudinal methods and technologies as well as in health behavior research. Physical activity (PA) serves as an important behavioral pathway to delayed functional decline, disease, and mortality as well as to improved quality of life and psychological well-being in older age. Yet, many people do not obtain the recommended levels of PA, with insufficient PA being more likely in older adults. Existing studies increasingly indicate that people who believe that low PA reflects an expected and inevitable part of the aging process report lower levels of PA engagement, but the pathways explaining this relationship are unclear. Whereas theories of aging suggest that negative self-views of aging (NS-VOA) adopted earlier in life become internalized and self-fulling later in life, health behavior theories emphasize the lack of motivation to engage in PA and poor self-regulation of PA behavior as driving forces. Guided by these ideas, the proposed research uses an intensive within-person approach to characterize the relationship and underlying mechanisms between PA and NS-VOA as they unfold in daily life. K99 Years 1 and 2 will use existing data to address Aims 1 and 2. Aim 1 will characterize the extent of within-person variation in NS-VOA, testing whether NS-VOA vary significantly on four timescales: years, weeks, days, and moments (i.e., within-days). Aim 2 will examine the association between NS-VOA and PA on daily and momentary timescales. It is hypothesized that PA engagement will be lower on occasions when NS-VOA are higher than usual, with stronger relationships for higher (vs. lower) intensity PA and when device-based (vs. self- reported) PA are used. Aims 1 and 2 are novel, within-person assessment contexts important for understanding fast-acting relationships and underlying mechanisms. R00 Year 3 will collect and analyze new data for Aim 3 to test whether NS-VOA act through motivation and self-regulation factors to influence PA on daily and momentary timescales. This work will inform preparation and submission of an R-grant (R00 Years 4 and 5) that focuses on prevention and intervention to increase older adults’ PA at key times when NS-VOA may impair PA engagement.

NIH Research Projects · FY 2025 · 2021-06

PROJECT SUMMARY / ABSTRACT Project SCORE Systemic inequities in access to healthcare and quality educational opportunity for the youth of Mississippi (MS) are powerful forces that disengage students from entering pathways toward STEM careers. Project SCORE (Student Centered Outcomes Research Experience) proposes to engage students from groups underrepresented in STEM career trajectories through a youth participatory action research (YPAR) approach, capturing and retaining their attention through the immediacy of health threats such as COVID-19. Guided by a YPAR approach, Project SCORE will leverage a near-peer mentor model to engage and support underrepresented students in an exploration of the field of public health. Project SCORE will bring together underrepresented high school and graduate health sciences students from two communities with significant health disparities in a year-long weekly afterschool program to develop relevant health behavior research questions, provide training in research methods, and facilitate the development of student-conducted research projects mentored by near-peer graduate health sciences students and faculty. A student-centered research agenda will be developed to support future research initiatives. Students will complete their experience with a week-long on campus immersion experience. This project seeks to increase awareness of and interest in public health, science engagement, and STEM careers, as well as increase matriculation into higher education STEM programs to enhance and diversify the future biomedical workforce.

NIH Research Projects · FY 2025 · 2020-08

PROJECT SUMMARY Pili are bacterial surface structures that are used in many ways. They can be used for motility, genetic exchange, and surface attachment, and are often critical virulence factors for pathogens. A particular type of pilus, the flp pilus, is found in many bacterial pathogens, including Vibrio vulnificus and Aggregatibacter actinomycetemcomitans, and is used by some members of the human microbiome to establish permanence in the intestinal tract. Despite its importance, the flp pilus is not nearly studied to the depth of other pilus systems, and has many unexplored facets and curiosities. In this proposal we outline experiments to explore the flp pilus using a model bacterium that provides many technical benefits for studying pili: Caulobacter crescentus. A common characteristic of the flp system is that the pilin gene is regulated separately from the rest of the pilus genes. In C. crescentus, the transcriptional activator CtrA binds to four sites in the pilin promoter. We have shown that one site induces expression, while the remaining sites inhibit and delay transcription from their upstream positions, a phenomenon not previously reported in bacteria. The inhibitory sites increase the affinity of CtrA for the promoter. We have also shown that relief of upstream inhibition leads to premature pilus production which results in increased susceptibility to a bacterial virus that uses the pilus to infect. Regulation of the pilin promoter will be explored by mapping CtrA binding over a concentration range in different promoter variations to identify which sites become occupied first. The spacing between binding sites and within the sites themselves will be investigated by altering spacing and testing for inhibitory activity through a variety of means. Lastly, we will explore the real-world consequences of premature pilus production by examining cell survival rates under persistent phage threat in a microfluidics chamber (Aim 1). Our previous work has identified an uncharacterized predicted RNase that appears to associate with the pilA promoter that inhibits transcription in some constructs but promotes expression in others. We will investigate the role of this RNase by examining its DNA binding ability, its role in RNA stability, and the proteins it interacts with (Aim 2). Lastly, we have shown that several different lineages of bacterial viruses infecting diverse Alphaproteobacteria convergently evolved usage of the host CtrA to control virus gene expression. We hypothesize that this is a Trojan horse mechanism to increase virus spread. We will investigate this hypothesis by characterizing the timing of virus gene expression during infection, disrupting CtrA regulation of virus genes by mutating the virus using novel methodology, and determining the impact of these mutations on gene expression (Aim 3). These studies will provide insight into unusual features of flp pili systems, and thus provide insight into a structure important for many human pathogenic and commensal organisms.

NIH Research Projects · FY 2024 · 2020-07

OVERALL: ABSTRACT/SUMMARY The University of Mississippi (UM) Botanical Dietary Supplements Research Center (BDSRC) is focused on filling in knowledge gaps related to the potential for the Spirulina-based product, ImmulinaTM, to promote resilience against and/or recovery from influenza and, by extension, other respiratory viral infections. ImmulinaTM is a patented bioassay-standardized extract that concentrates the active immunostimulatory compounds in Spirulina, i.e., Braun-type lipoproteins. Researchers affiliated with the UM BDSRC have generated evidence that supports reproducible and mechanistically plausible effects of ImmulinaTM for enhancing host responses that may promote resilience against respiratory viral infections such as influenza. Last year in the U.S. alone, influenza resulted in 959,000 patient hospitalizations and 79,400 deaths. Under the direction of Ikhlas Khan, PhD and Nirmal Pugh, PhD, the UM BDSRC includes a well- established collaborative team of multi-disciplinary investigators with expertise in natural products, animal models of human diseases, pharmaceutics, immunology, and human subject research. The UM BDSRC will be composed of an Administration Core, a Botanical Core, and two research projects: (1) Unraveling Immune Enhancement by ImmulinaTM, and (2) Evaluation of ImmulinaTM Oral Supplement for Host Resistance to Influenza Virus Infection. The overall specific aims are to: (1) maximize coordination of effort, communication among researchers, training opportunities for career development, and dissemination of knowledge through a well-managed and effective Administrative Core; (2) ensure product integrity of the ImmulinaTM extract through a combination of bioassay- and chemical-based approaches for standardization within the Botanical Core; (3) determine the molecular mechanism of immune enhancement of ImmulinaTM and optimal formulation and dosing in Project 1’s mouse study that can be utilized by the UM BDSRC’s other studies; and (4) using both a mouse and subsequently a biomarker-based human model, establish the impact of ImmulinaTM supplementation on increasing host resilience against the pathogenic effects of influenza virus infection with Project 2. The Administrative Core, guided by an External Advisory Committee and an Internal Steering Committee, will oversee all aspects of the UM BDSRC, including progress towards meeting its milestones, efficient allocation of resources, collaborations with researchers outside the UM BDSRC, oversight of its graduate students and post-doctoral fellows, and maintenance of its website. Members of the Botanical Core will work closely with researchers from the UM BDSRC to provide sufficient quantities of standardized ImmulinaTM extract and to collaborate with other units of CARBON on future efficacy studies of ImmulinaTM. The results from Projects 1 and 2 will help establish the optimal regimen for future clinical efficacy trials.

NIH Research Projects · FY 2025 · 2020-05

Project Summary The Glycoscience Center of Research Excellence (GlyCORE) was established in 2020 to develop a regional hub of glycoscience excellence in the IDeA state-rich Mid-South region. Glycoscience is an essential and rapidly emerging field of biomedical science and the challenging nature of glycoscience research requires tools and expertise that are not commonly found in the biomedical research community. Moreover, the biomedical community is rapidly growing to appreciate the essential role of carbohydrates, with investigators outside of the dedicated glycoscience community facing challenging questions in glycoscience as part of the course of their biomedical research. In Phase 1, GlyCORE established three Research Cores covering essential services to support modern glycoscience: the Analytical and Biophysical Chemistry Research Core, the Imaging Research Core, and the Computational Chemistry and Bioinformatics Research Core. GlyCORE also instituted a faculty development program including recruitment of new glycoscience faculty, financial support of the investigators' research efforts, a formal mentoring program, and priority subsidized access to University of Mississippi Core facilities through both a Research Project program and a smaller Pilot Project Program. Finally, GlyCORE initiated efforts to establish the University of Mississippi as a regional center for glycoscience in the Mid-South, including hosting an annual Mid-South Glycoscience Meeting and a monthly Seminar Series to highlight the role of glycoscience at the University of Mississippi and bringing in leaders in the glycoscience field. In Phase 2, we will build upon our progress in Phase 1, refine our approach to improve metrics for success, and prepare for our transition to long-term sustainability. We will expand the services offered by our Research Cores, including expanding our workshop offerings to broaden our impact and user base. We will support four concurrent Research Project Investigators throughout Phase 2 to improve the competitiveness of our researchers for major extramural funding, as well as supporting a total of nine one-year Pilot Projects to develop research ideas from their initial stages and aid in new faculty recruitment. In Phase 2, we will institute a cost-recovery model as part of our transition to sustainability, using a voucher system to provide full Core support to our internal glycoscience investigators. We will institute new programs to improve our investigator extramural funding success rate through an enhanced external mentoring program and incorporation of professional external grant proposal review services for current and previous Project Leaders. By the end of Phase 2, GlyCORE will be recognized as a major destination for glycoscience in the Mid-South, with a core of faculty expertise and sustainable research infrastructure necessary to support modern glycoscience.

NIH Research Projects · FY 2025 · 2018-09

PROJECT SUMMARY/ABSTRACT Hydrogenases are complex metalloenzymes that generate energy for certain organisms or balance cellular redox potentials by catalyzing the reversible interconversion between H2 oxidation and H+ reduction, respectively. Unraveling the intricate details of the function of these enzymes will significantly advance H2- based, carbon-neutral alternative energy production. However, the complexity of these enzymes due to the presence of multiple metallic cofactors, low production yield, and deactivation makes studying these enzymes challenging. Our goal is to design new biomolecular artificial hydrogenases (ArHs) as functional analogs of these complex metalloenzymes. De novo design and repurposing of native proteins are appealing and well-established approaches to creating active sites of complex metalloproteins within minimal protein scaffolds. Although the designed systems are less complex, they serve as water-soluble functional analogs of the native metalloenzymes and provide a functional view of the chemistry. Employing these approaches, we propose to pursue two Specific Aims describing the overall design principles and functional/mechanistic attributes of the ArHs inspired by the [NiFe] hydrogenases. The overall objectives of this proposal are: i) to design mononuclear, multinuclear, and heterometallic active sites within designed biomolecular scaffolds; ii) characterize the physical and catalytic properties of these ArHs; iii) determine the timescales of electron transfer; iv) tune the pKa of ligand and outline the role of ligand protonation states in H2 evolution; v) identify the metallated species responsible for noncooperative H2 production; and vi) elucidate how multimetallic sites influence the properties/reactivity, ultimately attaining a wholesome view of H-H bond formation. Our strong preliminary results presented here attest that our objectives are achievable. Collectively, the results from this proposed work will impact the fields of metalloprotein design, bioinorganic chemistry, and alternative energy research. A novel class of ArHs will emerge, which will provide functional vignettes into the working principles of H+ reduction related to the native enzymes. The lessons from this study will enable us to prepare biosynthetic catalysts with novel properties and functions in the future.

NIH Research Projects · FY 2025 · 2018-06

PROJECT SUMMARY/ABSTRACT Structural biology plays a central role in modern molecular bioscience, enabling both a greater understanding and new mechanisms of manipulation of biomolecular action. However, despite tremendous development in tools for the generation of high resolution molecular models, large families of biomolecules and biomolecular complexes are still poorly represented in databases of protein structure due to limitations of current technology, and methods for probing protein structure within mammalian tissue are few. One method that has been used successfully to qualitatively study the structure of several of these families is hydroxyl radical protein footprinting (HRPF), an emerging technology that has been used to study changes in protein topography by measuring changes in the apparent rate of reaction between hydroxyl radicals generated in situ and amino acid side chains on the protein surface. Our initial work has developed HRPF into a quantitative measurement of protein topography at the individual amino acid level, accurately measuring the average solvent accessible surface areas (<SASA>) of many individual amino acids in a single experiment. In this renewal, we will expand our technology into structural systems that change dynamically with time, including protein posttranslational modification systems, large heteromeric protein complexes, and protein:carbohydrate complexes. The core technology we will develop to enable these studies is high performance liquid chromatography coupled inline with amino acid resolution HRPF (LC-HR-HRPF). Inline liquid chromatography allows the separation of protein conformers and immediate quantitative measurement of the purified conformers’ topographies by HR-HRPF before the dynamic system has a chance to re-equilibrate, freezing the structural information in the stable chemical footprint. We will also develop technology for analysis of protein structure within mammalian whole blood, enabling the study of protein structure and interactions within highly complex native systems. We will develop flow systems to precisely and carefully deliver hydrogen peroxide to blood for protein labeling without damaging cells, and will demonstrate the technology with the structural analysis of monoclonal antibodies dosed into a mouse model. Finally, we will develop technologies to probe the topography of complex carbohydrates, enabling us to measure which parts of carbohydrates mediate interactions with proteins, even in complex mixtures of glycans. We will develop both reducing-end specific and non-specific labeling strategies for probing carbohydrate topography. Together, these advances represent potential transforming technologies for the structural analysis of biomedically important and highly challenging systems.