The phase I effort produced an effective agrochemical screening technique using tomato leaf and brain cell cultures, quantitative fluorimetry and specific fluorescent probes. A robot produced highly uniform cultures for screening four marketed agrochemicals; Alar, Atrazine, Malathion and Metrifonate. Chemical effects were monitored for commercially important crop and the mammalian end user. Phase I fluorimetirc assays measured compound toxicity, cell metabolism, protease activity and detected changes in neurotransmitter receptor signaling following 1 and 24 Hr. exposure to agrochemicals. In Phase II the applicant proposes to construct two patented In Vitro Fluoro-test kits; one for plant cells and one for mammalian cells. The Fluoro-test kit represents a reproducible, simple means of transferring this technology to companies developing and screening agrochemicals. This technology is not currently available. Continuation in Phase II will include development of an insect Fluoro-test kit. Initial studies on insect tissues are reported as having been fruitful. Larval cell culture and a semi-intact preparation will also be used to screen for novel pesticides in venom libraries derived from Cone snails. These natural products (venom peptides) target specific invertebrate channels and receptors without effecting vertebrate targets. Peptides should offer a novel approach to developing new agrochemicals. Sequences of these peptides will act as patterns for small molecule chemistry.
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