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Group Research Interests - an overview. James F. Rusling Professor of Chemistry and Pharmacology University of Connecticut and University Health Center. Example 1. Toxicity Screening. Lipophilic Molecule. Cyt P450, O 2. Enzyme-activated molecule. + DNA. Damaged DNA.

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Group Research Interests - an overview

James F. Rusling

Professor of Chemistry and Pharmacology

University of Connecticut and University Health Center


Example 1. Toxicity Screening

Lipophilic Molecule

Cyt P450, O2

Enzyme-activated molecule

+DNA

Damaged

DNA

Detect by voltammetry, LC-MS/MS


Collaboration with Prof. John Schenkman,

Pharmacology, Uconn Health Center

Funding from NIH, NIEHS


Screening Chemical Toxicity

Enzyme reaction - Incubate:

Reactant + H2O2-->metabolite

Analysis by catalytic SWV or

electrochemiluminescence

RuL2+ = RuL3+ + e-

RuL3+ + DNA-G --> RuL2+ + DNA-G•


Enzyme/DNA films

Peak increase

measures damage

of DNA by enzyme-

generated

metabolite



Comparison of toxicity sensors with LC-MS

For DNA damage by methylmethane sulfonate




Arrays detect in-vitro DNA damage from metabolites of Benzo[a]pyrene Metabolites

different enzymes with DNA/enzyme films

Rel. turnover rate,

1/min (nmol Enzyme)

Drug development

applications


Sensors for oxidative stress via oxidized DNA Benzo[a]pyrene Metabolites

Os-PVP

PSS

Pyrolytic Graphite

Ru-PVP

ECL detection in films:

Lynn Dennany, Robert J. Forster, Blanaid White, Malcolm Smyth and James F. Rusling, J. Am. Chem. Soc., 2004, 126, 8835-8841.


Single-Walled Carbon Nanotube Benzo[a]pyrene Metabolites

Forests: Antigen-Antibody Sensing

SPAN or

Nafion

Chattopadhyay, Galeska, Papadimitrakopoulos, J. Am. Chem. Soc. 2001, 123, 9451.

End COOH groups allow chemical attachment to proteins (antibodies)


(b) Benzo[a]pyrene Metabolites

(a)

AFM of SWNT forest with and without antibody attached

Collaboration with F. Papadimitrakopoulos (ARO funded)

SWNT + antibody

(EDC coupling)

SWNT


H Benzo[a]pyrene Metabolites2O2

HRP

HRP

HRP

HRP

HRP

HRP

Ag

Ag

Ab1

Ab1

Sandwich Assay for Human Serum Albumin

Ab2

HPR

SWNT forest

Apply E measure I


Detection of Human Serum albumin in Benzo[a]pyrene Metabolites

10 mL drops on SWNT forest immunosensor

LOD ~ 10 pmol/mL


Styrene Epoxidation Catalyzed by Benzo[a]pyrene Metabolites

Myoglobin-Polyion Films

in Microemulsions (NSF funded)

Stable in microemulsions



Additional Current Projects Benzo[a]pyrene Metabolites

• “Green” chiral synthesis using enzyme films in microemulsions, nanoparticle catalysts (NSF)

• Redox chemistry of complex photosynthetic proteins

in membrane films (with H. Frank, USDA funded)

• detection of Lyme disease vectors (proteins) in tick saliva (with Uconn Health Center)

• enzyme activity for generating toxic metabolites (with Uconn Health Center)

• EC-Laser fluorescent detection of DNA damage

• arrays for anticancer biomarkers (NIH)

• molecular dynamics of film construction


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