SBIR-STTR Award

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.

Thesaurus Terms:
biotechnology, environmental toxicology, fluorimetry, method development, neurotoxicology, pesticide biological effect, tissue /cell culture antifungal agent, bioassay, herbicide, pesticide brain cell, fluorescence microscopy, fluorescent dye /probe, immunocytochemistry, mass tissue /cell culture, plant

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.

Thesaurus Terms:
biotechnology, environmental toxicology, fluorimetry, method development, pesticide biological effect, tissue /cell culture, toxicant screening animal poison, biological signal transduction, computer program /software, insecticide biological effect, ion transport, metamorphosis, neurotoxicology, neurotransmitter receptor, pesticide, plant physiology, robotics bioengineering /biomedical engineering, brain cell, fluorescent dye /probe, larva, mass tissue /cell culture, organ culture, plan