Medical Chronobiology Laboratory



		Chronbiology and Oncology Research Laboratory Dr. William J.M. Hrushesky, MD Dr. Patricia A.Wood, MD, PhD
		The Medical Chronobiology Laboratory is a medium-sized NCI- and VA-funded laboratory dedicated to the study and compartmentalization of time-dependent biological variability into rhythmically predictable and residual realms. The laboratory resides within the WJB Dorn VA Medical Center at Columbia, South Carolina. Dr. William Hrushesky and Dr. Patricia Wood represent its professional staff, who are complemented with between ten and fifteen part-time and full-time technical staff, undergraduate, graduate, medical post-graduate students and visiting fellows throughout the world working under the auspices of either the University of South Carolina, School of Medicine or School of Public Health. Strong collaborations have also been forged between this laboratory and laboratories in University of South Carolina and the Dorn Research Institute, as well as a large number of biomedical device, biotechnology, and pharmaceutical firms. These and other relationships provide regular opportunities to impact developmental diagnostics and therapeutics and rapidly test or implement ideas within and outside of classical academic networks. The focus of the ongoing research in this laboratory is the effect of biologic rhythms upon host cancer balance, tumor biology, and the efficacy and toxicity of cytotoxic drugs and growth factors used in hematology and oncology. The biologic rhythms of interest include both circadian (~24 hour, daily), fertility (estrous, menstrual) cycle and seasonal rhythms. This studies include the use of both normal and tumor bearing humans and mice. This laboratory offers exposure to experimental design involving patients and whole animal as wells as in vitro cellular, biochemical and molecular studies. Weekly laboratory meetings are held where all participants present ongoing data and problems, share knowledge and techniques, and plan future studies.
		Specific projects include: Determining how core circadian clock (and clock controlled) genes coordinate cellular proliferation in normal tissues and cancer. Using core circadian clock gene knockout mice and tumor lines to understand how clock genes gate essential functions. Studying the effects of circadian disruption by light on core circadian clock gene expression and circadian gating of cancer cell proliferation and growth. Understanding how the female reproductive cycle phase at breast cancer resection impacts metastatic cancer spread and breast cancer outcome (cure). The following is general outline of some of the basic techniques utilized in this laboratory: Experimental Design Chronobiologic design, strategies Experimental animal handling Chronobiologic procedures Serial endpoints- temperature, weight, WBC/differentials, HCT, RTC, estrous, serum collections Tissue procurements Surgery - tumor resections, device implants, tumor or drug treatments and techniques Slide staining and reading WBC differentials Estrous stages Tissue preparations for biochemical analysis including sonication, centrifugation Clonal assays of hematopoietic progenitor cell types Enzyme Assays- Thymidylate Synthase, other Chemical Analyses, protein growth factor quantitations by immunoassays and biologic assays Protein Gel Electrophoresis SDS-PAGE gels Western Blots - membrane based, antibody detection, and quantitation of specific proteins Computer Skills Word processing - experimental protocol, methods Spread sheets - data collection and data storage/entery Graphics - data representation Data Analysis - Repeated Measures Analysis, tests, ANOVA, linear regression, Cosinor Analysis, Chi Squared Analysis, Life Table/Kruskall Wallis Analysis Computer Languages include - Java, C++, Visual Basic.Net, HTML, COBOL, SQL, JavaScript Technical Support (Computer and Internet) On-site equipment includes: -Computers: Six PC Computers with (4) 2.3Gig processors and (2) 1.2Gig processors Windows 2000 Network server Two Macintosh PowerComputing PowerCenter 150s (21" and 17" monitors) Two Macintosh II computers (15" and 14" monitors) Two Macintosh Quadra 800s (16" monitors) Three Macintosh SEs Macintosh Color Classic Northgate 386 Relisys Infiniti Scorpio scanner (graphic) Various laserprinters, including an HP LaserJet 4 and an NEC SilentWriter -Lab: (~1330 sq. Feet) 2 biologic flow hoods 2 infrared Forma CO2 37ºC incubators IEC, Forma and Sorvall refrigerated centrifuges 3 balances, 2 microcentrifuges (one temp. controlled) 3 American Optic light, 1 Nikon light and one Nikon phase microscopes 2 refrigerators, one -20ºC freezer One Forma Ultra Low -80ºC freezor 1-D gel electrophoresis setups (3) Gel dryer, transblot, water baths pH meters, shakers Spectrophotometer, chemical hoods, Skatron automatic plate harvester, pipets and pippetors, drying oven Large volume refrigerated centrifuges and refrigerated microfuges Mettler and digital top loading balances Torque electronic time switch for controlling 4 separate light staggered animal rooms Animal procedure rooms Access to common -80ºC freezer, Packard liquid scintillation and gamma counter, automatic spectrophotometric plate reader, Beckman and Sorvall ultracentrifuges, Molecular Dynamics computerized densitometer with computer, common tissue culture facility with 2 biologic flow hoods and 3 CO2 incubators, fluorescent microscopes, distilled/deionized water, autoclaves, dark room and film developer, and a full engineering service. We also have our own secretarial support with one full time secretary in our office. There is also a full service library and medical media department in the building. In summary, this laboratory is a totally unique, intense and exciting operation networking many fields through an unusual biological perspective and represents a real opportunity to impact medicine through a number of disparate avenues.