Capabilities

In vitro Models

• Precision –cut Tissue slices in Dynamic Organ Culture.  Suitable for comparative studies utilizing: liver, lung, and/or kidney slices from humans and laboratory animals.
• Primary tissue explant culture models utilizing human endometrium.
• Primary culture of isolated hepatocytes and lymphocytes.
• Cell culture utilizing various cell lines.

Biochemical/Molecular Techniques

• Morphological and biochemical assays for cellular viability.
• Xenobiotic uptake and metabolism.
• Assay of chemical-DNA adducts by ³²P post-labeling.
• Cell and tissue specific expression of cytochrome P-450s (CYPs):
  
  o Immunohistochemical location.
  o Quantitative Competitive RT-PCR analysis of specific mRNA.
  o Western immunoblotting.
  o CYP enzymatic activity.

Toxicogenomics

• Single Stranded Conformation Polymorphism (SSCP) analysis
• Restriction Fragment Length Polymorphism (RFLP) analysis
• Site directed mutagenesis.

Bioassays

• Reporter Gene Bioassay for dioxin-like compounds (96 well plate format).

Preclinical Toxicology Testing

The Center has protocols for the preclinical testing of drugs in support of New Drug Applications.  The primary focus has been on new radiodiagnostic drugs, particularly brain imaging agents.  Examples of publications in this area are provided below.

Kostyniak, P.J., Brenner, J.S., MacCubbin, A.E., Nakeeb, S., Kasliwal, R., and Kung, H.F.:  The toxicity and mutagenicity of S-3-Iodo-N-(1-ethyl-2-pyrrolidinyl) methyl-2-hydroxy-6-methoxy benzamide (IBZM), a new NS D-2 dopamine receptor imaging agent. Drug and Chem. Toxicol., 11(4):  433-442 (1988).

Kostyniak, P.J., MacCubbin, A.E., Nakeeb, S.M., John, E.K., Green, M.A., Schopp, E.M. and Kung, H.F.: Acute Toxicity and Mutagenicity of the Copper Complex of Pyruvaldehyde-bis-(N-4-methylthiosemicarbazone), Cu-PTSM. J. Appl. Toxicol. 10: (6), 417-421 (1990).

Equipment

• Krumdieck precision cut tissue slicer
• 2 Custom designed incubators for dynamic organ culture, sonicator, electrophoresis and blotting apparatus
• Perkin Elmer 9600 GeneAmp PCR system
• Avanti J-251 centrifuge
• Beckman L-70 Ultracentrifuge
• PTI Spectrofluorometer
• Wallac 1409 liquid scintillation counter
• Dynatech MRX-HD micro plate reader
• CytoFluor II fluorescent plate reader
• Bio-Rad gel doc 1000 fluorescent gel documentation system
• Bio-Rad model Gs-525 molecular imager system
• Bio-Rad GS-700 densitometer
• PT1 Delta scan dual scanning fluorometer
• Class II type B-2 biological safety cabinets

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