?url_ver=Z39.88-2004&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Adc&rft.relation=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F6882%2F&rft.title=A+Cytopathic+Effect-Based+High-Throughput+Screening+Assay+Identified+Two+Novel+Compounds+that+Inhibit+Dengue+Infection%3A+Streptovitacin+A+and+Nagilactone+C&rft.creator=McCormick%2C+Kevin+Dylan&rft.description=Dengue+is+an+emerging+infectious+disease+and+is+spreading+world-wide+at+exponential+levels.+Two+billion+people+in+over+100+countries+are+at+risk+for+infection+from+one+of+the+four+serotypes+of+the+dengue+virus.+Those+infected+with+dengue+may+develop+diseases+such+as+dengue+fever+and+dengue+hemorrhagic+fever+(DHF)+and+of+the+500%2C000+cases+that+progress+to+DHF+each+year%2C+more+than+22%2C000+will+result+in+fatality.+Discovering+new+antivirals+to+treat+DHF+is+essential+to+reducing+this+disease+burden.+Here%2C+we+have+developed+a+cytopathic+effect-based+high-throughput+screen+(HTS)+to+discover+possible+inhibitors+of+Dengue+viral+infection+of+hepatocytes+in+vitro.+Dengue+virus+infection+of+hepatocytes+induces+massive+cell+death%2C+%22cytopathic+effect+(CPE)%22%2C+which+we+converted+into+a+screening+assay+whereby+inhibitors+of+Dengue+infection+prevent+cells+from+dying.+In+this+assay%2C+the+viral+induced+CPE+is+quantitated+by+monitoring+cellular+ATP+levels%2C+which+positively+correlates+with+cellular+viability.+ATP+in+the+cell+culture+will+drive+the+oxidation+of+luciferin+resulting+in+the+emission+of+light+that+is+quantitated+using+a+luminometer.+The+assay+is+simple+and+highly+reproducible+yielding+a+screening+window+coefficient%2C+Z-factor%2C+of+0.78%C2%B10.12+between+plates.+The+Z-factor+is+a+statistical+parameter+commonly+accepted+as+an+assay+quality+assessment+and+is+reported+as+a+value+0+to+1+and+anything+over+0.5+is+considered+excellent+quality.+This+assay+is+advantageous+to+current+methodology+as+it+simultaneously+screens+possible+inhibitory+compounds+while+controlling+for+any+unwanted+toxicity+triggered+by+these+drugs.+Our+initial+HTS+of+a+288+small+compound+library+yielded+a+total+of+eleven+hits+that+prevented+the+CPE+of+dengue+infection.+Further+evaluation+with+an+immunofluorescence+assay+showed+that+two+of+these+compounds%2C+Streptovitacin+A+and+Nagilactone+C%2C+are+highly+potent+inhibitors+of+dengue+infection.+At+effective+inhibitory+doses%2C+they+did+not+appear+to+be+cytotoxic%2C+and+therefore+both+of+these+compounds+are+possible+antivirals+and+could+be+used+to+elucidate+various+cellular+mechanisms+utilized+during+the+dengue+life+cycle.+The+discovery+of+these+two+inhibitors+demonstrates+the+efficacy+of+our+newly+developed+assay+and+the+public+health+significance+of+this+project.&rft.date=2011-06-29&rft.type=University+of+Pittsburgh+ETD&rft.type=PeerReviewed&rft.format=application%2Fpdf&rft.language=en&rft.identifier=http%3A%2F%2Fd-scholarship-dev.library.pitt.edu%2F6882%2F1%2Fmccormickkd_etd2011.pdf&rft.identifier=++McCormick%2C+Kevin+Dylan++(2011)+A+Cytopathic+Effect-Based+High-Throughput+Screening+Assay+Identified+Two+Novel+Compounds+that+Inhibit+Dengue+Infection%3A+Streptovitacin+A+and+Nagilactone+C.++Master's+Thesis%2C+University+of+Pittsburgh.++++(Unpublished)++