RESOURCES AND ENVIRONMENT

Dr. Mahato’s laboratories are located in the Cancer Research Building, University of Tennessee Health Science Center, 19 Manassas, Memphis, TN 38163. Dr Mahato’s laboratories are equipped with most of the modern equipment necessary for conducting research in delivery systems of proteins, oligonucleotides, siRNA and genes. These laboratories house inverted light and fluorescence microscopes, Chemifluorescent and UV Gel Imaging System, two Thermal Cyclers for PCR, several Gel electrophoresis apparatus, UV transilluminator,Western blotting apparatus, two microcentrifuges, bench-top centrifuge, two UV visible Spectrophotometers, two cell culture hoods, three incubators that can control levels of N₂ and air, in addition to CO₂, two -30°C freezers, one -80°C ultrafreezer, two refrigerators, and cuvette washer, rotary incubator. We also have rotary evaporator, two chemical hoods, vacuum aspirator, pH meters, microbalance, hot air ovenS, vacuum oven, CAMAG UV housing system, freeze-dry system and JULABO water bath. We have Malvern Nano ZS which can measure particle size, zeta potential and molecular weight. We have recently purchased an Waters Breeze^TM HPLC/GPC systems for purification, quantification and molecular weight measurement of polymers and bioconjugates. Our Department of Pharmaceutical Sciences has core facilities of HPLC-MS, FT-IR, UV-Vis Spectrophotometer, NMR and Molecular modeling. We have full access to the 30,000 sq ft Animal Resources Division (ARD).

Dr. Mahato’s group has access to the 4C cold room, dark room for film development available in the Cancer Research Bldg. We also have access to other laboratories of the Department of Pharmaceutical Sciences in Crowe and Johnson Buildings and Islet Isolation Laboratory in Coleman’s Building. These are modern laboratories, with all the equipment needed for polymer synthesis, purification and characterization, gene cloning, plasmid amplification and purification, formulation, in vitro and in vivo characterization of polymer/DNA complexes, animal surgery, cell culture, islet transplantion, microscopy, cold rooms. UT Health Science Center has several core facilities having many modern equipment including DNA micro-array, DNA sequencing, DNA and Peptide Synthesis, Flow Cytometry and Quantitative PCR.

Core Facilities at the University of Tennessee Health Science Center:

Molecular Resource Center: The Molecular Resource Center (MRC) is located in the 3^rd floor of the Cancer Research Bldg, 19 Manassas, Memphis, TN 38163. The MRC maintains satellite laboratories in eight other locations within the UTHSC campus partnering with the Departments of Comparative Medicine, Pharmacology, Anatomy and Neurobiology, Neurology, Pediatrics and Molecular Sciences. MRC houses genotyping (Applied Biosystems 3700 and 3100 DNA Analyzers), sequencing (Applied Biosystems 310 and 3100 Sequencing Analyzers), LightCycler®480 new realtime PCR, Real Time PCR Applied Biosystems 7700, and Primer Express Software to determine primers and probes for real time PCR), flow cytometry (phenotyping, DNA analysis, cell sorting, and phosphor imaging (chemiluminescent, radioactivity, fluorescence, Molecular Diagnostics Typhoon) equipment and employs a staff of six specialists. The MRC has recently acquired 3 single capillary genetic analyzers, high capacity 96 capillary gene mapping systems, 2 high capacity 16 capillary DNA sequencers, 2 DNS chip arrayers, DNA chip reader (fluorescent), 2 phosphorimager/flurimagers, and a microinjection system for transgenic production. New cytometric equipment includes a fluorescence activated cell sorter and laser capture microdissection microscope. The LightCycler® 480 System setup enables the use of all current probe formats (e.g., SYBR Green I, Hydrolysis Probes, HybProbe Probes) and provides the ideal solution for fast and precise qualitative or quantitative detection of nucleic acids, genotyping, and mutation analysis

Molecular Biology, Immunogenetics and Vector Laboratory: provides the capability to generate adenoviral vectors for use in our genetic therapy transduction studies. This core support laboratory performs numerous outcome measures for the assessment of metabolic function in the islet tissue through chromatographic and PCR analyses. Flow cytometric apoptosis studies and mRNA upregulation as measured by Real Time-PCR are conducted in this laboratory.

Surgical Immunology/Flow Cytometry Laboratory is a core support facility for the Transplant Division at U.T. Memphis. It provides diagnostic services for transplant recipients including monitoring of cyclosporine, and tacrolimus levels in our recipient population. Additionally pre and post transplant Flow Cytometric Crossmatching is performed to determine the levels of T and B cell sensitization of recipients to donor organs. This facility is ASHI accredited and maintains service availability 24 hours per day, seven days per week.

Vascular Biology Center includes ten laboratories with approximately 4,500 sq. ft. Shared research facilities include radioimmunoassay and ligand preparation, assay, molecular modeling, gel analysis, medical imaging, cell and organ culture, gene transfer, homologous recombination, gene targeting and a transgenic mouse core facility.

Center of Excellence in Genomic and Bioinformatics Center (CGB) was recently established with $10 million of funding from the University of Tennessee 21^st Century Initiative. Headed by neuroscientist Dan Goldowitz, this interdisciplinary center builds upon a strong base in functional genomics and bioinformatics. The Center consists of 6 physical Cores (DNA analysis, mouse transgenic & knockout, microarray, proteomics, mouse mutagenesis, and bioinformatics) providing equipment and technical expertise critical to the field of functional genomics.

Proteomic Center: The Proteomic laboratory (under the Direction of Dr. Desiderio) is located in the Johnson Building. having Matrix-assisted laser desorption/ionization time-of-flight Mass Spectrophotometer (MALDI-TOF MS), PerSeptives Biosystems Voyager DERP Workstation; Liquid-chromatograph electrospray, Quadrupole ion trap mass spectrometers, ThermoQuest LCQ^DECA and LCQ^DUO; ISCO capillary zone electrophoresis. LC-electrospray quadrapole ion trap mass spectrometers, 500mHz nuclear magnetic resonance systems, fluorescence spectrophotometer and polarizers.

In addition, we have access to several of the facilities at St. Jude Children’s Research Hospital and the University of Memphis.

Islet Isolation Laboratory We have access to the Islet Isolation Laboratory is housed in the Coleman Medical Building. It has been in existence since 1992 with an active pre-clinical and clinical islet transplant program. Our islet isolation facility is a part of the NCRR (National Centres of Competence in Research) as National Islet Cell Resource Network and we have full FDA approval to proceed with human islet transplants using our facility. They have performed over 280 islet isolations for preclinical and clinical trials. Laboratory equipment includes three dedicated incubators, 2 laminar flow hoods, isolator biocontainment system, 2 water baths, 2 refridgerated centrifuges, 1 microscope, ethylene oxide gas sterilizer, fraction collectors, biocontainment air system, chiller, cryofreezer, liquid nitrogen dewar, liquid handling pumps, wrist action shakers, refrigerator, freezer, and Cobe cell separator. The Dean of Research at the UTHSC has pledged additional space (4,200 sq. ft) for construction of an expanded GMP compliant facility for islet isolation.