IN VITRO FLUORO-TEST FOR ENVIRONMENTAL/AGROCHEMICALS

DESCRIPTION: (Adapted from the Applicant's Abstract). This proposal will develop an agrochemical discovery and safety screening technology using plant and brain cell cultures tested with quantitative fluorimetry and specific florescent probes. Cultures of plant and brain cells will be grown in a statistically uniform way to test chemical assays. The first specific aim of this phase I proposal is to use robots to develop uniform cultures of tomato leaves and rat cortical neurons in a 96 well culture plate for high throughput screening of a variety of agrochemicals. The second specific aim is to test the effects of these chemicals on secondary targets, such as commercial crops. The third specific aim is to test these chemicals on CNS cultures to monitor the toxicity that may be encountered through environmental exposure. The overall goal is to produce the assays and a set of standardized kits for the agrochemical products discovery, as well as neurotoxicity testing. $ = TOTAL AWARD AMTS & NOT LIMITED TO PORTION OF PROJECT RELATED TO SUBJECT OF SEARCH SUBPROJECT $ = TOTAL AWARD AMOUNT DIVIDED BY NUMBER OF SUBPROJECTS SOURCE: CRISP FORMAT F FY 97 LAST UPDATE 04-07-98 1QUERY 1536 ID SEARCH 06/01/98 PAGE 289 --PROJECT NUMBER......1 R43 ES08857-01 INVESTIGATOR NAME/ADDRESS FY 97 LEOPOLD, PETER E IRG/INTRAMURAL UNIT..ZRG5 PROTEIGENE, INC AWARD AMOUNT......... $85,591 44 MANNING RD BILLERICA, MA 01821 PERFORMING ORGANIZATION: PROTEIGENE, INC. TITLE RAPID IDENTIFICATION OF INFECTIOUS BACTERIA ABSTRACT: DESCRIPTION: (adapted from investigator's abstract) The problem of rapid identification of pathogenic bacteria in the environment remains unsolved. The CDC estimates that each year 4,000 Americans die, and millions become sick, from poultry and meat tainted by bacteria, such as E. coli or Salmonella. Recent E. coli outbreaks in Japan, and at Jack-in-the-Box restaurants, show that lives could be saved if rapid identification was possible, in order to: (1) eliminate the pathogen source, and (2) take appropriate medical countermeasures. Recently, the Army's ERDEC published that MALDI-TOF-MS of either proteinaceous extracts, or of entire cells, has a high potential for specific identification of bacteria feared as biowarfare agents. Genus-, species-, and strain- specific protein biomarkers in the MALDI-TOF spectra were reported for Bacillus anthracis, Yersinia pestis, and Brucella melitensis. MALDI-TOF-MS measurements per se take less than 1 minute. Thus, bacteria identification can be fast provided the sample preparation protocol is rapid. ProteiGene, Inc. will research MALDI-TOF-MS for rapid protein biomarker identification of pathogenic bacteria - in Phase I particularly E. coli. ProteiGene will determine the optimal sample preparation and measurement conditions for obtaining reliable biomarker identification, and determine a pattern of MALDI-TOF-MS peaks which uniquely identify E. coli, and distinguish it from other bacterial species.