This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The COBRE Technical Support Core provides specific services and optimizes protocols to facilitate the successful implementation and completion of the COBRE Research Projects. The COBRE Mentor, Dr. Vivek R. Nerurkar oversees the utilization of core services and has overall responsibility for managing COBRE technical support resources of this Core. A part-time Molecular Biologist, one full time Interviewer/Research Assistant, and BSL-3 laboratory manager, a part-time Biostatistician, and a virology Research Assistant, staff the COBRE Technical Support Core. The Core consists of three principal technical support services: namely, a molecular biology and immunology core facility, a biostatistics and bioinformatics core facility, and a BSL-3 containment facility. Molecular Biology and Immunology Core Facility - A flow cytometer, purchased with NINDS equipment funds in September 2001 provides a hitherto unavailable capacity to the proposed project on the T-cell immune responses to dengue virus infection. The Core also optimizes real-time PCR protocols for quantitation of viral load assays and of gene expression. The core assists COBRE Investigators in standardizing protocols for microarray analysis and for confirming the differentially expressed genes using real-time PCR and Western blot. Biostatistics and Bioinformatics Core Facility - The COBRE biostatistician, Dr. Wang Kit Cheung, assists Projects 1 and 3 PI, and pilot project 2 PI in planning their experiments. Similarly, the biostatistician conducts didactic workshops for junior investigators in statistical methods. Dr. Jimmy Efird, the newly hired biostatistician with RCMI support, also assists the COBRE investigators in conducting biostatistical analysis. Sequence and phylogenetic analyses are currently conducted by the Virologist within the COBRE Core. Dr. Shannon Bennett will join the COBRE Technical Core as a bioinformatician in summer 2004. With her strong background in evolutionary biology, specifically dengue virus evolution, she will be an asset to the COBRE. BSL-3 Containment Facility - The availability of a BSL-3 facility allows studies on dengue virus, West Nile virus (WNV) and on other agents of medical importance in the Asia-Pacific region. Plaque neutralization assays were conducted on both strains of West Nile viruses, NY99 and Egypt 101, and the virus was used in the studies conducted by Pilot Project 2 PI. Similarly, the BSL-3 laboratory is used for conducting neutralization assays for dengue virus. The COBRE supported BSL-3 facility will serve as a fulcrum in the future development of the new Pediatric Dengue Vaccine Initiative and to build the Arbovirus Reference Center at the University of Hawaii. To support the State of Hawaii Department of Health (SOH-DOH) project entitled 'Public Health Preparedness and Response for Bioterrorism' funded by the CDC, the SOH-DOH and the University of Hawaii have signed a contract to build a new BSL-3 suite next to the current BSL-3 suite. The new 250 sq. ft. BSL-3 suite will be constructed and fully equipped by monies allocated by the CDC to SOH-DOH. This new facility is anticipated to be functional by end of summer 2004 and will also serve the needs of COBRE Technical Core. With the rejuvenation of the Department of Tropical Medicine and Medical Microbiology (TMMM) at the University of Hawaii, and hiring of a world class arbovirologist Dr. Duane Gubler, to establish the Asia Pacific Center for Tropical Medicine and Medical Microbiology, the COBRE supported Technical Core will be involved in supporting the new tenure-track faculty who are anticipated to join the Department of TMMM by December 2004. Equipment: Apart from a BSL-3 suite and radiation facility, the COBRE investigators have full access to the following equipment: FACSCalibur Flow Cytometer, Phosphorimager, Top Count Microplate Scintillation and Luminescence Counter, Fluorimeter, PCR machines, Real-Time Thermal Cycler (purchased using COBRE funds), Ultracentrifuge (floor and table-top models), Speed Vac, Sorvall High-Speed Refrigerated Centrifuge, Fume Hood, PCR and cloning, and sequencing related equipment. The COBRE Technical Core works closely with the University of Hawaii Molecular Biology Core Facility (UHMBCF), which is based on a charge-back system. Major UHMBC equipment used by the COBRE investigators are: ABI PRISM 377 DNA sequencer, Affymetrix Gene Chip DNA Microarray System including Array Maker and Scanner and BioAnalyzer.
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