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Biomedical subjects

Bingquan Wang

Publications and source records attributed to Bingquan Wang.

6 recordsLinked to original sources

Genotoxicity sensor response correlated with DNA nucleobase damage rates measured by LC-MS.

Responses from "reagentless" DNA-based electrochemical toxicity sensors to DNA alkylating agents styrene oxide (SO), diepoxybutane (DEB), and methyl methanesulfonate (MMS) were compared to formation rates of total alkylated nucleobases in DNA measured by LC-UV-MS. Sensors utilized a catalytic metallopolymer in DNA films previously exposed to the damage agents. To achieve adequate sensitivity, LC-UV-MS analyses were done on DNA in solution reacted with the damage agents, and subsequently hydrolyzed to nucleosides with enzymes. Sensor response correlated well with nucleobase-adduct formation rates obtained by the molecule-specific analyses. Results confirm that the metallopolymer-DNA film sensors can be used to estimate relative DNA damage rates from nucleobase adduct-forming chemicals. Results from both methods correlated well with animal genotoxicity as estimated by TDL(o) values, the lowest dose producing carcinogenicity, in mice and rats. These sensors should be useful for rapid, inexpensive screening of moderately and severely genotoxic new chemicals.

Animals↗

Evaluating enzymes that generate genotoxic benzo[a]pyrene metabolites using sensor arrays.

Arrays with individually addressable, demountable electrodes coated with ultrathin DNA/enzyme films were evaluated to estimate relative rates of genotoxic bioactivation of benzo[a]pyrene (BP) for several different enzymes simultaneously. Specifically, cytochrome (cyt) P450cam, cyt P40 1A2, and myoglobin in the array were activated with H2O2 to metabolize BP to genotoxic metabolites. DNA damage by the metabolites was detected by increases in square wave voltammetric oxidation peaks using Ru(bpy)3(2+) as catalyst. Cyt P450cam and cyt P450 1A2 showed 3-fold higher activity for genotoxic bioactivation of BP than myoglobin. The ability of the arrays to generate and detect metabolite-based DNA damage simultaneously for several enzymes is a rapid and promising approach to identify and characterize enzymes involved in genotoxicity of drugs and pollutants.

7,8-Dihydro-7,8-dihydroxybenzo(a)pyrene 9,10-oxide↗

Voltammetric sensor for oxidized DNA using ultrathin films of osmium and ruthenium metallopolymers.

Films containing [Os(bpy)2(PVP)10Cl]+ and [Ru(bpy)2(PVP)10Cl]+ metallopolymers were assembled layer by layer on pyrolytic graphite electrodes to make sensors that selectively detect oxidized DNA. These films showed reversible, independent electrochemistry for electroactive Os3+/Os2+ and Ru3+/Ru2+ centers, with formal potentials of 0.34 and 0.76 V vs SCE, respectively. The combination of ruthenium and osmium metallopolymers in the films provided a catalytic Os square wave voltammetry (SWV) peak that is mainly selective for 8-oxoguanine and the detection of other oxidized nucleobases from the Ru peak. The method is applicable to measurements on DNA in solution or DNA incorporated into films. Using the Os SWV peak, 1 oxidized nucleobase in 6000 was detected. The sensor is simple and inexpensive, and the approach may be useful for the detection of oxidized DNA as a clinical biomarker for oxidative stress.

Biosensing Techniques↗

Voltammetric sensor for chemical toxicity using [Ru(bpy)2poly(4-vinylpyridine)10Cl)+ as catalyst in ultrathin films. DNA damage from methylating agents and an enzyme-generated epoxide.

Films containing presynthesized [Ru(bpy)2poly(4-vinylpyridine)10Cl)]Cl and ds-DNA grown layer by layer by alternate electrostatic assembly were used to detect DNA damage from an epoxide metabolite and methylating agents on a reaction time scale of minutes. The redox polymer [Ru(bpy)2poly(4-vinylpyridine)10Cl)]Cl was used as an inner layer in films 14-25 nm thick to catalyze the voltammetric oxidation of guanine bases of ds-DNA in the outer layers. This film architecture provides a self-contained, reagentless sensor for toxicity screening based on detection of DNA damage. Films were incubated with reactants and washed, and then DNA damage was analyzed by square wave voltammetry (SWV). Bioactivation of styrene to its metabolite styrene oxide was accomplished by incorporating the protein myoglobin into the films to catalyze the conversion. DNA damage caused the catalytic SWV peaks at approximately 0.75 V vs SCE to increase nearly linearly over the first 10-20 min of reaction, depending on the damage agent employed. Such prototype toxicity biosensors hold promise for in vitro screening of new agricultural chemicals and drugs for potential genotoxicity.

2,2'-Dipyridyl↗

Lipid membrane immobilized horseradish peroxidase biosensor for amperometric determination of hydrogen peroxide.

Stable films of didodecyldimethylammonium bromide (DDAB, a synthetic lipid) and horseradish peroxidase (HRP) were made by casting the mixture of the aqueous vesicle of DDAB and HRP onto the glassy carbon (GC) electrode. The direct electron transfer between electrode and HRP immobilized in lipid film has been demonstrated. The lipid films were used to supply a biological environment resembling biomembrane on the surface of the electrode. A pair of redox peaks attributed to the direct redox reaction of HRP were observed in the phosphate buffer solution (pH 5.5). The cathodic peak current increased dramatically while anodic peak decreased by addition of small amount H(2)O(2). The pH effect on amperometric response to H(2)O(2) was studied. The biosensor also exhibited fast response (5 s), good stability and reproducibility.

Biosensing Techniques↗

A method to construct a third-generation horseradish peroxidase biosensor: self-assembling gold nanoparticles to three-dimensional sol-gel network.

A novel method for fabrication of horseradish peroxidase biosensor has been developed by self-assembling gold nanoparticles to a thiol-containing sol-gel network. A cleaned gold electrode was first immersed in a hydrolyzed (3-mercaptopropyl)-trimethoxysilane (MPS) sol-gel solution to assemble three-dimensional silica gel, and then gold nanoparticles were chemisorbed onto the thiol groups of the sol-gel network. Finally, horseradish peroxidase (HRP) was adsorbed onto the surface of the gold nanoparticles. The distribution of gold nanoparticles and HRP was examined by atomic force microscopy (AFM). The immobilized horseradish peroxidase exhibited direct electrochemical behavior toward the reduction of hydrogen peroxide. The performance and factors influencing the performance of the resulting biosensor were studied in detail. The resulting biosensor exhibited fast amperometric response (2.5 s) to H2O2. The detection limit of the biosensor was 2.0 micromol L(-1), and the linear range was from 5.0 micromol L(-1) to 10.0 mmol L(-1). Moreover, the studied biosensor exhibited high sensitivity, good reproducibility, and long-term stability.

Biosensing Techniques↗