• Dissecting Protein and Nucleic Acid Contributions to Efficient tRNA Aminoacylation
  Awarded $100,000 for the period 10/1/07-9/30/09
  Source: National Foundation for Cancer Research

As essential participants in protein biosynthesis, the aminoacyl-tRNA synthetases (ARSs) are potential targets for novel anti-infective agents, particularly for cancer patients whose immune systems are suppressed due to chemotherapy. This project investigates the long-standing question of how ARSs transmit a binding event to an amino acid attachment site that can be tens of Ångstroms away. High-resolution structures of ARSs bound to their substrates often reveal conformational changes in the protein and/or tRNA. Mutually induced fit between protein and tRNA may lower the activation energy barrier to catalysis, as in the “action at a distance” seen in allosteric enzymes. Computational, spectroscopic, and structural methods will be used to characterize the conformational rearrangements that occur upon ligand binding in both a model protein and its homolog from a pathogenic organism.

• CAREER: Dissecting Domain/Domain Communication in Methionyl-tRNA Synthetase
  Awarded $123,974 for the period 6/1/08 to 5/31/09
  Source: NSF

• Support for Research Technician for Collaborative Projects
  Awarded $36,000 for the period 5/2/05 to 5/1/06
  Source: Scripps Research Institute

  The Scripps Research Institute has offered to pay the salary of a research technician to facilitate three collaborative projects with Dr. Alexander’s laboratory. These structural and mechanistic studies of methionyl-tRNA synthetase (MetRS) extend and enrich Dr. Alexander’s current effort, supported by the National Foundation for Cancer Research.

• Biochemistry through Biotechnology: Undergraduate Laboratory Enhancement
  Awarded $18,000 for the period 8/1/05 to 7/31/06
  Source: NCBC

Funds will be used to purchase equipment and supplies to enhance the biotechnology training of WFU undergraduates. A new, one-semester biochemistry laboratory course taken at the junior or senior level, in association with the two-semester biochemistry lecture series developed jointly by the Chemistry and Biology departments, consists of an 11-week study of the enzyme lactate dehydrogenase (LDH), which students will isolate from beef heart, purify by chromatography, and quantify and assay using spectrophotometric methods. Molecular biology approaches, such as PCR of the LDH gene and transgenic expression, will also be taught. With the faculty now in place to expand the undergraduate curriculum to include biochemistry courses and a new local and regional emphasis on expanding biotechnology, WFU is poised to provide undergraduates the background to enter this growing employment market or to receive further academic training.

• Structural and Mechanistic Studies of Methionyl-tRNA Synthetase
  Awarded $25,000 for the period 4/1/05 to 10/31/05
  Source: National Foundation for Cancer Research

Dr. Alexander’s laboratory is pursuing three projects in collaboration with researchers at the Scripps Research Institute. The partners will undertake structural and mechanistic studies of methionyl-tRNA synthetase (MetRS) that will inform the development of cancer therapies.

• Research Infrastructure in Minority Institutions (RIMI)
  Awarded $7,716 for the period 10/1/06 to 9/29/07
  Source: National Institutes of Health (NIH)

When the temperature drops, bacteria significantly overproduce a protein called CsdA. It may work to unwind RNA, because at low temperatures, an increase in RNA secondary structure would inhibit mRNA translation at the ribosome. In collaboration with PI Pamela Jones, a microbiologist at Winston-Salem State University, Dr. Alexander will purify CsdA mutants and assay their effect on RNA binding and unwinding in vitro.

Ulrich Bierbach

• with Bruce King, Cemistry, Fred Salsbury, Physics, and Roy Hantgan, Biochemistry
  Molecules to Medicines: Crafting a New Interdisciplinary Curriculum in Drug Discovery
  Awarded $7,500 for the period 3/18/08 to 3/19/09
  Source: Camille and Henry Dreyfus Foundation

Sparked by keen student interest, Wake Forest’s mandate for entrepreneurship, and the pharmaceutical  industry’s need for cross-disciplinary scientists with critical-thinking and teamwork skills, the project aims to prepare students in the biological, chemical, and physical sciences to pursue wider career paths by developing a new course, Drug Discovery, Design, and Development: Molecules to Medicines, for advanced undergraduate and graduate students. Dreyfus funds will allow promising students in the course, Dreyfus Discoverers, to complete internships in the pharmaceutical and biotechnology industries. They will return to serve as teaching assistants, sharing their knowledge and experiences through seminars and weekly sessions where they will lead discussions of problems assigned by course faculty.

• Novel DNA-metalating Hybrid Anticancer Agents
  Awarded $220,019 for the period 5/1/12 to 4/30/13
  Source: NIH

This research uses a unique chemical approach to develop a new, platinum-based cancer treatment to manage intractable tumors, especially nonsmall cell lung carcinomas (NSCLC).

Christa Colyer

• Capillary electrophoresis in clinical chemistry
  Awarded $41,902 for the period 8/1/2012 to 7/31/13
  Source: Ameritox

Toxicological analysis of human fluids depends on separation technology for precise identification and quantification of drugs and drug metabolites. Recently, it has relied almost exclusively on liquid chromatography (HPLC or UPLC) coupled to a sensitive detector (e.g., mass spectroscopy, laser-induced fluorescence). While the resolution of these techniques is very good, capillary electrophoresis is better, without the attendant costs of columns, solvents, and pumps. This project aims to develop an effective quantitative CE/MS method for screening drugs of interest for pain medication monitoring.

• Acquisition of a User-accessible Q-TOF Mass Spectrometer
  Awarded for $442,102 for the period 1/1/10 to 12/31/12
  Source: NSF

The Department of Chemistry secured funds to purchase an Agilent 6530 Accurate-Mass Quadrupole Time-of-Flight (Q-TOF) mass spectrometer that will be available for hands-on use by faculty, postdocs, and students in a new Biomolecular Chemical Analysis Facility (BCAF). Its high sensitivity and high mass accuracy is crucial to metabolomics, unambiguously sequencing peptides, and analyzing posttranslational modifications of proteins. The new facility represents significant investment in a US-based company, Agilent Technologies, helping to preserve and create jobs, and our emphasis on student and postdoc training will help to expand the regional and national biotechnology workforce.

• Affinity-based CE Studies to Facilitate Bioprobe Design and Microbe Detection
  Awarded $113,000 for the period 7/1/10 to 6/30/11
  Source: NSF

Analytical tools must be able to deliver high-efficiency, high-sensitivity measurements for a wide range of analyte types and sizes, especially targets relevant to human and environmental health. Capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) exploits noncovalent interactions between protein-based analytes and new fluorescent probes. This project aims to optimize analyte/probe couples for given applications; in particular, ligand/receptor interactions essential to the function of the B7 family of proteins, which is involved in regulating immune responses. It will also develop rapid, CE-based assays targeting residues unique to the surfaces of analytes important to human health and security, such as the influenza virus and anthrax spores. Finally, it will create holistic CE-LIF methods to analyze intact organisms, their unique and characteristic constituent proteins, and important submolecular pigments. Broader impacts include a summer research program for High Point University undergraduate students; an annual Electrophoresis Working Weekend for faculty and students from regional undergraduate colleges and universities; an international research student exchange with Osaka Prefecture University in Japan; and a series of case studies, Separation Shorts, designed around modern separation science and biosensing methods, to help college instructors develop their students’ problem solving.

• Noncovalent labeling and CE-LIF strategies for the determination of intact microorganisms
  Awarded $75,000 for the period 8/1/07 to 1/31/09
  Source: NCBC

Analytical tools that can be adapted to a wide range of analyte types are invaluable in interdisciplinary science. They must deliver measurements with high efficiency and high sensitivity, especially for targets within complex matrices. Two strategies involve noncovalent interactions between protein-based analytes and new fluorescent probes that use capillary electrophoresis with laser-induced fluorescence detection (CE-LIF). First, however, their governing physicochemical parameters must be characterized and then important, innovative applications explored. The project aims to devise tools and methodologies for supramolecular entities, such as intact microbes.

Successful completion of these studies should facilitate development of reagent kits and mobile, point-of-contact CE-LIF instruments capable of simple, inexpensive, and rapid fingerprinting of a variety of microorganisms with applications in the fields of forensics, homeland security, clinical screening, and environmental protection.           

• Collaborative Exploration: Using CE to Quantify PE Pigment Concentrations
  Awarded $5,000 for the period 8/1/06 to 7/1/07
  Source: National Aeronautics and Space Administration (NASA)

This project aims to demonstrate the utility of capillary electrophoresis with laser-induced fluorescence detection for analyzing more phycobiliproteins than those previously extracted from a single synechococcus culture. By applying the group’s extraction and analysis protocols to 15 different samples, including other synechococcus cultures, a cryptomonad culture, and a rhodamonas culture, the project will evaluate the comparative effectiveness of this methodology and begin the process of modification. 

• Real-time Bioaerosol Sensor with Airborne Taggant Applicator
  Awarded $80,000 for the period 8/25/05 to 8/24/06
  Source: Department of Homeland Security (DHS)

The project will develop a low-cost, real-time, optical bioaerosol sensor that can discriminate many types of naturally occurring biological aerosols from biological aerosol threats and improve the lower detection limit for singlet spores through the airborne application of two optical taggants. Airborne particles are illuminated one at a time in two separate but correlated regions that allow five measurements.

The resulting sensor will significantly enhance an existing bioaerosol trigger. It is expected to deliver a response in less than one minute with a false-alarm rate of less than once per year with a 98% confidence level at an agent concentration of 1000 CFU/liter of air. The system is expected to detect all classes of agent: spore, cellular, viral, and protein toxin. The sensor is also expected to come close to the desired cost characteristics with low annual maintenance costs.

• with Thomas A. Arcury, Family and Community Medicine
  Cadmium, Lead, and Arsenic Exposure among North Carolina Farmworkers
  Awarded $20,000; $10,000 Reynolda campus, $10,000 Health Sciences
  Source: WFU Cross-Campus Collaborative Research Fund

  The project seeks to identify the risks in working at farm sites previously treated with toxic metal-containing pesticides and fertilizers. While organic pesticides are known risks to migrant workers, the risks from exposure to metal-containing agricultural products are not well understood. Farmworkers will be recruited, complete questionnaires, and provide urine samples. The project will develop a method to determine cadmium, lead, and arsenic levels in urine and, once perfected, used to analyze 260 samples. The toxic metal body burden will be compared with other potential risk factors associated with farmworker lifestyle, and the results disseminated to the at-risk population.
  

• A Small Portable Tungsten Coil Spectrometer
  Awarded $5,000 for the period 11/18/05 to 11/17/06
  Source: WFU Science Research Fund

  A low-cost, portable atomic spectrometer will be designed and evaluated for use in field applications, such as environmental monitoring and drinking water testing. It will use a tungsten coil as an atomization device and to generate continuum radiation. The instrument will measure either atomic absorption or atomic emission spectra, depending on the optical alignment. The atomizer will be coupled to a small, hand-held, high-resolution detection system using a fiber-optic cable. The system will be the simplest and most portable multi-element atomic spectrometer ever described.
• Faculty Recruitment Grant: Assistant Professor of Chemistry
  Awarded: $100,000 for the period 6/1/05 to 5/31/06
  Source: NCBC

Funds will assist in recruiting a third Assistant Professor of Biochemistry (1) to complete the biochemistry division on the Reynolda Campus; (2) to encourage recruitment of undergraduate and graduate students in the program; and (3) to enhance biological expertise in western North Carolina. The successful candidate will help to develop the curricula in biological chemistry and teach courses leading to the BS and BA degrees in chemistry with a concentration in biochemistry. The candidate will also establish a research program that complements current faculty efforts. The purchase of major equipment and supplies will support the university's bioscience infrastructure as a whole, and Wake Forest will match grant funds with start-up funding and by renovating biochemistry research space.

Paul Jones

• Oxidative Radical Cyclizations: Mechanistic and Synthetic Investigations
  Awarded $233,700 for the period 9/1/05 to 8/31/08
  Source: NSF

  Dr. Jones’s laboratory has discovered a new photochemical reaction of anthraquinoes. It consists of two unprecedented chemical steps: the cyclization of a phenol radical on a tethered alkene and electron transfer from a carbocation to a semiquionone radical. The project will first establish the reaction’s scope and synthetic utility, with the hope of using it to prepare biologically active anthraquinones. Next, it will try to extending the reaction’s scope beyond anthraquinones to any phenol with an alkene tethered to the ortho position. Results could have repercussions for physical organic as well as synthetic organic chemistry.

• Ionic Lubricants Incorporating Nanomaterials
  Awarded: $31,000 for the period 9/30/04 to 6/29/05
  Source: Air Force Office of Scientific Research (AFOSR)

  The project teams Dr. Jones’s lab with NanoTechLabs to synthesize and to evaluate ionic liquid/carbon nanotube-based lubricants to reduce bearing and other mechanical wear. They aim, first, to use unique cations, based on chiral imidazoliums developed at Wake Forest, as a basis for the lubricant. The presence of chiral centers in the alkyl substituents in combination with chiral anions in a salt depresses its melting point and prevents crystallization. Commercially available anions will be combined with the new cations to form the lubricant. Second, they will incorporate single-walled carbon nanotubes (SWNTs) into the ionic lubricants to reduce friction and to enhance thermal stability. Carbon nanotubes should affect the molecular ordering of the lubricant and change with the lengths of the nanotubes.

Mechanical properties will be evaluated by pin-on-disk tribometer. Nanoscale wear analysis (nanotribology) using atomic force microscopy (AFM) will evaluate lubricant film thickness and morphology as well as any boundary layer formed by lubricant decomposition. AFM will allow film properties to be measured through a range of temperatures by use of hot and cold stages. Scanning electron microscopy (SEM) will permit microstructural analysis of the wear surface, and nanotube length and dispersion will be characterized by transmission electron microscopy (TEM).

Currently, lubrication in space mechanisms, electronics, and the computer industry concentrates on perfluoropolyethers (PFPEs), which, however, have poor solubility and are susceptible to autocatalytic degradation. Ionic liquids have a number of desirable characteristics, including negligible volatility, nonflammability, high thermal stability, low melting point, and broad liquid range. Versatile lubricants of use on a variety of materials, such as ceramics, aluminum, and steel, will be of great value to the Air Force and industry. Because of their controlled miscibility with organic compounds and low volatility, ionic liquids may also play an important role in green synthesis.

• New Photochemistry for Visible Light-Initiated Photoactivation of Enzymes and Biocides
  Awarded $215,530 for the period 4/1/04 to 3/31/07
  Source: NIH

Photodynamic Therapy (PDT) treats diseases with light-activated chemotherapeutics and most often involves the formation of singlet oxygen via energy transfer from dye sensitizers. Singlet oxygen is highly reactive and relatively unselective, and the sensitizer can make the patient sensitive to bright light for weeks.

The long-term goal of this project is to develop a new PDT system that does not use singlet oxygen or leave the patient sensitive to light. Its achievement will require a new chemistry and biochemistry for the photochemical production of biologically active molecules.

Specific aims are:
1) To develop a system that can act as a photolabile, amine-protecting group using visible light;
2) To photoactivate enzymes in vitro;
3) To develop photochemistry capable of photoreleasing biologically active aldehydes; and
4) To demonstrate in vitro biological activity that depends on visible light.

These aims are based on the hypothesis that photoactive agents with more selective biological activity will prove more predictable and allow the tailoring of PDT agents to specific diseases. For example, a photoactivated clot-busting enzyme could form the basis of an effective treatment for Central Retinal Vein Occlusion, which currently has no treatment, while a photoactivated antiviral agent could treat a number of dermal viral infections. An effective general photoactivation scheme for biomolecules could have limitless therapeutic applications.

Angela Glisan King

• Project SEARCH Academy Summer Follow-up Program in Chemistry
  Awarded $3,902 for the period 6/18/12 to 6/22/12
  Source: North Carolina Area Health Education Centers (AHEC)

Project SEARCH Academy is a one-week program designed to increase student interest in biomedical careers by preparing them for rigorous high school science classes. The 2012 program focuses on chemistry.

• Project SEARCH
  Awarded $3,954 for the period 6/1/11 to 6/24/12
  Source: Wake Forest Baptist Health

Project SEARCH Academy’s Summer Follow-Up Program in Chemistry is designed to prepare students for high school chemistry and, ultimately, science careers through laboratory experiences. Participants are introduced to medical applications of concepts and the quantitative rigor required for success in chemistry.

• with Leah H. McCoy, Education
  WINS = Wake Innovative Noyce Scholars
  Awarded $893,753 for the period 7/1/09 to 6/30/14
  Source: NSF

Wake Forest University’s Noyce Scholars Phase 1 project will recruit, train, and mentor through induction 32 teachers highly qualified to lead grade 9-12 biology, chemistry, mathematics, and physics classes. Undergraduate STEM majors and professionals will be aggressively recruited, seeking a diverse cohort in disciplinary and demographic terms. The 13-month program includes extensive study of the teaching and learning process based on best practice research, leading to licensure in a STEM field and a Master’s degree in Education. Graduates will be required to teach for two years in a high-need school while formally mentored through documented interactions with the Advisory Board and co-PIs. Postgraduation mentoring mechanisms include: (1) email, discussion boards, and blogs; (2) two annual on-campus seminars; and (3) financial support to attend professional conferences. A Project Evaluator will examine WINS operations, effectiveness, and impact from quantitative and qualitative data including surveys, teaching videos, teaching artifacts, and student achievement scores. As a result, annual cadres of committed, innovative, and effective STEM teachers will maximize the achievement of students in high-need schools. Project results, including analysis, conclusions, and reflections, will be disseminated in presentations and publications to develop a national model for improving teacher education and retention in STEM fields.

• Rural Exposure to Biotechnology
  Awarded $4,925 for the period 5/15/04 to 7/1/05
  Source: NCBC

This partnership between Wake Forest University and South Stokes High School aims to prepare and to encourage a greater number of high school students from the rural target audience to pursue interests and careers in biotechnology at any level. This objective will be achieved by providing: electrophoresis and enzyme kinetics equipment and lab supplies not used in standard high school experiments and thus not provided by the school budget at a rural North Carolina high school; training for all the school's science teachers on how to use the equipment and ideas for incorporating biotechnology into the Standard Course of Study; and a field trip for the rural students to a biotechnology laboratory to broaden their career horizons.

• SCIMAX
  Awarded $82,263 for the period 9/1/05 to 8/31/06
  Source: NSF

SCIMAX (SCIENCE AND MATH EXCELLENCE) is a community-driven, K-16 partnership to ensure that all students graduating from Winston-Salem/Forsyth County Schools are able to pursue postsecondary studies and/or careers in science and mathematics.

S. Bruce King

• Synthesis of 33177 as an influenza antigen
  Awarded $7,056 for the period 11/7/12 to 5/31/13
  Source: NIH/Wake Forest Baptist Health (WFBH)

The R848 adjuvant, which is not commercially available, will be produced in a form that can be conjugated to influenza particles in the vaccine for the studies in a funded application

• Ca093389-targeted nanoparticles for kidney cancer therapy
  Awarded $25,000 for the period 9/15/11 to 9/14/12
  Source: Army Research Office (ARO)/WFBH

Dr. King will contribute the chemistry expertise necessary to ligate D5s to nanoparticles.

• Development and Evaluation of Acyloxy Nitroso Compounds as Nitroxyl Donors
  Awarded $42,805 for the period 9/1/10 to 9/1/11
  Source: Cardioxyl Pharmaceuticals, Inc.

Acyloxy nitroso compounds form a mechanistically distinct group of nitroxyl (HNO) donors that require the removal of the acyl group to release HNO. Variation in acyl group structure changes the ease of hydrolysis and controls HNO release in simple acyloxy nitroso compounds. While reactivity with esterases provides rapid HNO formation, it prevents long-lasting acyloxy nitroso-based HNO donors from developing. This project aims to lengthen their lifetime.

• Proteomic profiling of cancer-related redox signaling pathways
  Awarded $10,000 for the period 5/1/09 to 4/30/10
  Source: NIH

Reactive oxygen species (ROS) play a major role in carcinogenesis and many cancer therapies, such as ionizing radiation, cisplatin, and taxanes. Recently, cancer cells have been shown to produce ROS. This project has developed new experimental and computational technologies to identify the molecular targets modified as a result of either ROS damage or ROS signaling. This novel labeling technology will be developed: 1) to investigate the basic mechanisms of ROS damage and signaling; 2) to stratify patients with cancers that are sensitive to ROS-generating therapies via molecular profiling; and 3) to develop novel cancer therapies based on inhibiting ROS-dependent proliferative signaling.

• Nanotubes in tumor imaging and therapy
  Awarded: $53,612 for the period 8/1/09 to 7/31/11
  Source: NIH/WFU Health Sciences (WFUHS)

Carbon nanotubes have unique properties, including the ability to generate heat when stimulated with infrared light. This project will fabricate carbon-based nanotubes of different lengths, architectures, and composition to assess their capability as antitumor agents in tissue culture and mouse models. Mathematical modeling will optimize their placement for heat delivery and antitumor effect with minimal damage to adjacent tissues. The ultimate goal is a precisely directed therapy with low nonspecific toxicity that is compatible with standard clinical imaging instruments and easily cleared by the body.

• Niroxyl/Nitric Oxide-producing reactions of hydroxyurea and related compounds
  Awarded $330,126 for the period 7/6/10 to 6/30/11
  Source: NIH

Hydroxyurea has been approved for treatment of sickle cell disease, and exciting work reveals important roles for nitroxyl (HNO) and nitric oxide (NO) in sickle cell pathophysiology and treatment. Dr. King’s laboratory has identified C-nitroso species and HNO as important components of NO-formation from hydroxyurea. Developing new HNO/NO donors based on C-nitroso compounds will define structural entities with unique cardiovascular properties. The project aims to determine 1) the kinetics, reaction products, structural requirements, and extent of enzyme activation/inhibition in reactions of HNO derived from C-nitroso compounds with the heme-containing proteins catalase, soluble guanylate cyclase, and cytochrome P450; 2) whether C-nitroso compounds act as NO/HNO donors; and 3) the interaction of these new NO/HNO donors with biological target molecules and tissues, including hemoglobin, soluble guanylate cyclase, catalase, peroxisomal proliferator-activated receptor gamma (PPARy), pre-constricted blood vessels, and cardiac tissue. Results will advance the use of HNO/NO donors to treat sickle cell disease, other anemias, and congestive heart failure.

• Proteomic Profiling of Cancer-Related Redox Signaling Pathways
  Awarded $10,000 for the period 5/1/09 to 4/30/10
  Source: NIH

Reactive oxygen species (ROS) play a major role in carcinogenesis and many cancer therapies, such as ionizing radiation, cisplatin, and taxanes. While cancer cells have been recently shown to produce ROS as signaling molecules that promote proliferation, the molecular details of are far from clear. This project has developed new experimental and computational technologies uniquely suited to identifying the molecular targets that are modified by ROS, as a result of either ROS damage or ROS signaling. Its novel labeling technology will be developed for application to: 1) investigation of the basic mechanisms of ROS damage and ROS signaling; 2) molecular profiling to stratify patients with cancers that are sensitive to ROS-generating therapies; and 3) development of novel cancer therapies based on the inhibition of ROS-dependent proliferative signaling.

• Profiling of Redox-Sensitive Signaling Proteins
  Awarded $5,331.64 for the period 5/1/07 to 4/30/09
  Source: NIH/WFUHS

For over twenty years, redox mechanisms have been implicated in carcinogenesis, but the lack of large-scale methods to identify proteins that respond to cellular redox changes is a serious barrier to progress. This project hypothesizes that redox signaling affects the initiation of cell proliferation and transformation. Because the tools to test it directly are not available, Dr. King and collaborators will integrate analytical protein chemistry, cell biology, and bio-informatics to develop the reagents and methods to identify modifications in proteins involved in signal transduction pathways. Successful development of this technology will allow the underlying hypothesis to be tested in large-scale future research.

• Synthesis of Hydroxamic Acids through NOH Insertion of Ketones
  Awarded $50,000 for the period 7/1/08 to 8/31/09
  Source: Petroleum Research Fund

The project’s long-term goal is to develop a new method to prepare N-substituted hydroxamic acids by inserting NOH into ketones. Hyroxamic acids play many roles in a variety of biological systems containing metals, especially siderophores, molecules that mediate iron acquisition and uptake in numerous micro-organisms. Current methods to prepare N-substituted/cyclic hydroxamic acids rely on chemistry that is complicated by both N and O nucleophiles. This project will approach the synthesis of substituted hydroxamic acids through the basic decomposition of benzenesulfohydroxamic acid (Piloty’s acid, or PA) in the presence of a ketone. Initial experiments will establish the mechanistic aspects of this unique reaction; then reaction conditions will be optimized and the scope of this reaction sequence explored. Finally, these synthetic transformations will be applied to synthesis of the naturally occurring cyclic hydroxamate cobactin, found in the siderophores produced by the Mycobacteria, including Mycobacterium tuberculosis. Results will provide an alternative, efficient method for preparing these biologically important molecules.

• with Daniel B. Kim-Shapiro, Physics
  Effects of Nitric Oxide in Sickle Cell Blood
  Awarded $345,112 for the period 5/1/11 to 4/30/12
  Source: NIH

The project will elucidate the biochemistry and biophysics of nitric oxide (NO) in sickle cell blood and its use as a treatment for the disease. NO may benefit patients as a vasodilator, decreasing red blood cell sickling and sickle cell adherence and improving oxygen transport. However, much of NO biology in both sickle cell and normal blood is not well understood, and previous studies report contradictory results. Results from sickle blood will be compared to those in 1) normal blood; 2) preparations of isolated normal and sickle red blood cells; and 3) purified hemoglobins. A variety of spectroscopic and other biophysical tools, some developed specifically for this project, will include microscopy, ektacytometry, ultracentrifugation, stopped and quench-flow mixing, laser photolysis and diffraction, chemoluminescence, electron spin resonance, nuclear magnetic resonance, and absorption spectroscopy.

• with Ulrich Bierbach, Chemistry, Fred Salsbury, Physics, and Roy Hantgan, Biochemistry
  Molecules to Medicines: Crafting a New Interdisciplinary Curriculum in Drug Discovery
  Awarded $7,500 for the period 3/18/08 to 3/19/09
  Source: Camille and Henry Dreyfus Foundation

Sparked by keen student interest, Wake Forest’s mandate for entrepreneurship, and the pharmaceutical  industry’s need for cross-disciplinary scientists with critical-thinking and teamwork skills, the project aims to prepare students in the biological, chemical, and physical sciences to pursue wider career paths by developing a new course, Drug Discovery, Design, and Development: Molecules to Medicines, for advanced undergraduate and graduate students. Dreyfus funds will allow promising students in the course, Dreyfus Discoverers, to complete internships in the pharmaceutical and biotechnology industries. They will return to serve as teaching assistants, sharing their knowledge and experiences through seminars and weekly sessions where they will lead discussions of problems assigned by course faculty.

• with Daniel Kim-Shapiro, Physics
  Nitric Oxide Donor Compounds for the Treatment of Hemolytic Conditions
  Awarded $162,657 for the period 1/1/09 to 12/31/09
  Source: NIH

Nitric Oxide (NO) is synthesized in the endothelial cells surrounding blood vessels and signals smooth muscles to relax and increase blood flow. Hemoglobin (Hb) actively scavenges NO. In normal physiology, its scavenging is reduced due to  its compartmentalization in red blood cells, but in several diseased conditions, including hemolytic anemias, such as sickle cell disease and paroxysmal nocturnal hemoglobinuria (PNH), thalassemia intermedia, malaria, thrombotic thrombocytopenic purpura, hemolytic uremic syndrome, and cardiopulmonary bypass, Hb is released into the plasma compartment and can efficiently scavenge NO. This increased NO scavenging leads to a host of complications contributing to morbidity and mortality.

Administering NO through inhalation restores normal NO responsiveness and shows promise as a treatment, but it is not practical for chronic treatment. This project tests and, to some extent, synthesizes compounds that may eventually be taken intravenously or orally. Unlike many therapeutic compounds, they will be cell-impermeable, acting in the plasma compartment to react preferentially with cell-free Hb and inactivate its NO scavenging ability. Lead compounds include NONOates (NO donors), nitroxyl donors, and possibly nitrated lipids and nitrites.

• Nitric Oxide-Producing Reactions of Hydroxyurea
  Awarded $47,058 supplement for the period 12/1/06 to 11/30/07
  Source: NIH

Hydroxyurea is a new treatment for sickle cell disease, a painful condition that affects 1 in 600 Americans of African descent. The project's long-term goal is to explain how hydroxyurea's nitric oxide (NO)-producing reactions contribute to sickle cell therapy in order to improve it. Product analysis and kinetic and spectroscopic studies will determine the mechanism of hydroxyurea's NO production in vitro, and similar studies of various tissues and purified enzymes incubated with hydroxyurea will reveal both the mechanism and site of in vivo NO formation. Specifically, these results will distinguish hydrolytic from oxidative mechanisms.

Spectroscopic methods and product analysis will also be used to demonstrate the extent of enzyme activation and the ultimate products and kinetics of the reaction of hydroxyurea-derived NO and the identified target proteins, soluble guanylate cyclase and cell-free hemoglobin. These results will indicate the ability of hydroxyurea or hydroxyurea-derived NO to influence or to react with these target proteins, providing evidence for a mechanism for the beneficial effects of hydroxyurea treatment. Unique hydroxyurea-based NO delivery systems and water-soluable nitroxyl donors will be prepared through chemical synthesis. Their ability to release NO will be determined using spectroscopic and cyclic voltammetric studies, and these results will reveal the ability of these new systems to act as NO or nitroxyl donors.

• with Charles Morrow, Biochemistry
Structural Requirements of Nitrated Fatty Acids: Natural Cellular Signaling Agents and Nitric Oxide Donors
Awarded $20,000 for the period 5/06-5/07
Source: WFU Cross-Campus Collaborative Research Fund

Recently, nitrated fatty acids have been identified as common constituents of various biologically crucial cells. Under physiological conditions, they activate the nuclear transcription factor, peroxisomal proliferator-activated receptor (PPAR), and donate nitric oxide (NO). This project will examine the relationship of their structure, particularly the position of nitration, to these functions. Defined nitrated fatty acids will be prepared by chemical synthesis and evaluated for their ability to bind PPAR and to induce transcription. They will also be evaluated as NO donors by various analytical techniques. Results should clearly define the structural requirements for activating PPAR and NO release and how this pathway is involved in both normal and pathophysiology.

• with Jacquelyn Fetrow, Computer Science and Physics
  Profiling of Redox-Sensitive Signaling Proteins
  Awarded $32,340 for the period 5/1/06 to 4/30/07
  Source: NIH

For over twenty years, redox mechanisms have been implicated in cancer development, but the lack of large-scale methods to identify proteins that respond to cellular redox changes is a serious barrier to progress. This project hypothesizes that redox signaling affects the initiation of cell proliferation and transformation. Because the tools to test it directly are not available, Dr. King and collaborators will integrate analytical protein chemistry, cell biology, and bioinformatics to develop the reagents and methods to identify modifications in proteins involved in signal transduction pathways. The successful development of this technology will allow the underlying hypothesis to be tested in a future broad-scale research project.

• Bio-Organic Chemistry of N-Hydroxyureas and Related Compounds
  Awarded $60,000 for the period 1/00 to 12/05
  Source: Dreyfus Foundation

  The Henry Dreyfus Teacher-Scholar Award supports the teaching and research of outstanding young chemistry faculty. Dr. King's research on the roles of nitric oxide in biological systems, much of it carried out by undergraduates, has implications for new treatments of sickle cell disease and a wide array of other problems.

• Design and Self-Assembly of Cluster-Based Materials
  Awarded a $16,000 supplement for the period 1/1/06 to 12/31/06
  Source: NSF

• Inter-American Materials Collaboration: Hybrid Inorganic-Organic Materials
  Awarded $33,000 for the period 7/15/05 to 12/31/06
  Source: NSF, CONACYT

  Dr. Lachgar and Dr. Munoz, a colleague at the Autonomous University of Morelos (UAEM), Mexico, will combine their expertise in solid-state materials and metal organic preparation and characterization to develop a collaborative research and educational program. They will investigate novel hybrid inorganic-organic materials involving multifunctional ligands, octahedral and cubic metals, or inorganic clusters. The first year will involve the two PIs, three graduate students, and two undergraduate students, one from each group. The study is expected to provide a framework for the PIs to educate a number of graduate and undergraduate students in the interdisciplinary field of solid-state hybrid inorganic-organic materials, which involves a number of synthesis and characterization methods necessary to any well-trained materials chemist.

Akbar Salam

• 21st Century Centre of Excellence Guest Professor
  Awarded JPN YEN 500,000 for the period 12/8/05 to 1/8/06
  Source: Kyoto University, Japan

• Visiting Fellow, Institute for Theoretical Atomic, Molecular, and Optical Physics (ITAMP)
  Awarded $4,000 for the period 3/19/07 to 4/20/07
  Source: Harvard University, Department of Physics


• Wiley-International Journal of Quantum Chemistry Young Investigator Award
  $1,000 plus conference registration
  Source: University of Florida’s Quantum Theory Project; the Sanibel Symposium on Atomic, Molecular, Biophysical, and Condensed Matter Theory; and John Wiley & Sons, Inc.

Mark Welker

• Sequential Reactions of Main Group Element-Substituted Dienes
  Awarded $126,000 for the period 1/13/10 to 1/31/11
  Source: NSF

The Welker group has developed a novel method that provides the first ready access to relative and absolute cycloadduct stereochemistries and core structures with biological activities ranging from insect antifeedants to medicine. This project aims (1) to prepare 2-main group element-substituted 1,3-dienes to enhance ease of handling and storage; (2) to determine the scope and limitations of a new synthetic methodology, tandem Diels-Alder/cross-coupling reactions of main group-substituted dienes; (3) to perform sequential transmetallation/Diels-Alder/hydrolysis reactions and transmetallation/ Diels-Alder/cross-coupling reactions of main group-substituted dienes; (4) to affect sequential oxidative addition/Diels-Alder/transmetallation/reductive elimination reactions of 2-halogen-1,3-dienes; and (5) to use this methodology to construct biologically important core structure targets. Students who work on this project are broadly trained in synthetic organic and organometallic chemistry, and most have secured careers in academia or the pharmaceutical or specialty chemicals industries.

• Preparation and Evaluation of P13 Kinase Inhibitors for Treatment of Prostate Cancer
  Awarded $152,002 for the period 6/1/09 to 5/31/11
  Source: NIH/WFUHS

This project aims to demonstrate proof of the principle that tumor targeting by both prostate-specific antibodies and PSA-mediated activation is more effective than single targeting. Analysis of clinical samples shows that up to 60 percent of androgen-independent prostate cancers have active P13 kinase, an enzyme critical to their growth and survival, and that small-molecule inhibitors of P13K induce these cells’ death, showing promise as therapeutic agents, although they are highly toxic. This project will first generate inactive pro-drugs by attaching a peptide that prevents interaction with P13K. They should be activated only at tumor sites, where prostate specific antigen (PSA) concentration is high, and remain inactive in circulation. Second, these pro-drugs will be conjugated to J591 antibodies that recognize PSMA, a protein expressed on the surface of prostate cancer cells, which should increase pro-drug concentration in prostate tumors. While anti-PSMA antibodies are known to accumulate in liver and kidneys, which limits their use as a carrier of active toxins, the P13K pro-drugs should remain inactive in these organs since their PSA concentrations are much lower than in prostate tumors. Computer modeling is used to determine where on the P13K inhibitor to attach a linker that can be coupled with a PSA-cleavable peptide. If this dual approach shows positive results in a mouse model of metastic prostate cancer, it could be streamlined into clinical trials.

• Wake Forest University CRADLE Program Kick-off, Grants Writing Workshop
  Awarded $2,100 for the period 7/1/09 to 8/31/09
  Source: NCBC

The Creative Research Activities Development and Enrichment (CRADLE) initiative supports faculty seeking external sponsorship for multiyear research projects and creative activities. Fellows articulate a 5-year career plan that incorporates proven strategies for obtaining funding. The program begins with an intensive two-day grantwriting workshop led by Dr. David Bauer. One day focuses on how to find and win government grants, and the second on foundation and corporate funding.

• New Organosulfur Anticarcinogenic Enzyme Inducers
  Awarded $208,705 for the period 4/1/05 to 3/31/08
  Source: NIH

The project’s long-term goal is to produce nontoxic cancer chemopreventive agents. A comprehensive cancer treatment strategy will ultimately involve the use of small molecules for both the treatment and prevention of cancer, but to date, much more progress has been made in identifying small molecule antitumor agents than small molecule cancer prevention agents. The proposed work helps to close this gap.

Chemoprevention of cancer involves the use of chemical agents either to retard or to block carcinogenesis. These agents affect the metabolism of xenobiotic procarcinogens by inducing the enzymes that detoxify potential carcinogens. Typically, phase 1 of xenobiotic metabolism involves oxidative processes, and phase 2, redox or conjugation chemistry. This project will search for chemical agents of low toxicity that elevate phase 2, but not phase 1, enzymes as a cancer prevention strategy.

• Preparation and Tandem Reactions of Main Group Substituted Dienes
  Awarded: $360,000
  Source: NSF

  The Welker group has shown that certain dienes can be used to reverse the normal endo selectivity of Diels-Alder reactions and provide access to new cycloadduct stereoisomers in high yield and diastereoselectivity. Now, they propose to study tandem reactions of these new compounds. Boron, silicon, and aluminum-substituted dienes offer practical advantages over transition metal-substituted dienes in terms of cost, preparation, and disposal and are more amenable to catalytic chemistry and tandem reactions. This methodology can also access biologically significant core structures that have applications ranging from insect antifeedants to biomedical science.

Their work has been cited several hundred times as of 2004, and other groups have used their previously reported cycloadducts and methodology for making new, functionalized carbocyclic amino acids. Four senior scientists/postdoctoral fellows, 6 graduate students, and 13 undergraduate students have received training in synthetic organic and organometallic chemistry.