By Herbert S. Rosenkranz, Elena C. McCoy, Monica Anders, William T. Speck, David Bickers (auth.), Michael D. Waters, Stephen Nesnow, Joellen L. Huisingh, Shahbeg S. Sandhu, Larry Claxton (eds.)
vi Williamsburg, Virginia, February 21-23, 1978. This symposium was once subsidized through the U. S. Environmental safety service provider, place of work of power Minerals and undefined, Washington, DC, and workplace of overall healthiness and Ecological results, health and wellbeing results Re seek Laboratory, Biochemistry department, examine Triangle Park, NC. The symposium consisted of 24 formal displays that magnify the 3 significant issues mentioned in the course of the symposium: an summary of momentary bioassay platforms; present method concerning the gathering and chemical research of environmental samples; and present study in volving using temporary bioassays within the fractionation and research of complicated environmental combos. the aim of this symposium was once to give the state of the art tech niques in bioassay and chemical research as utilized to com plex combos and to foster endured development of this crucial zone. complicated combinations mentioned comprise ambient air and water, waste water, ingesting water, shale oil, syn thetic fuels, car exhaust, diesel particulate, coal fly ash, cigarette smoke condensates, and foodstuff items. it really is our desire that this quantity will function a refer ence to catalyze and inspire additional study during this box. Michael D. Waters, Ph. D. Stephen Nesnow, Ph. D. vii Acknowledgment we wish to thank Gerald Rausa, place of work of strength Minerals and undefined, for his suggestion, encouragement, and aid of this application. we might additionally wish to convey our appreciation to Wendy A. Martin, Peter A. Murphy, and David F. Wright of Kappa structures, Inc.
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Additional info for Application of Short-Term Bioassays in the Fractionation and Analysis of Complex Environmental Mixtures
The dishes are stained with Giemsa 6-8 days later to determine cloning efficiency, and stained 14-16 days later to determine the frequency of ouabain-resistant colonies. The mutation frequency for resistance to ouabain is calculated per 10 6 survivors, based on cloning efficiency and number of cells seeded for mutant selection. The mutagenic activities of 11 hydrocarbons with different degrees of carcinogenicity were tested in the fibroblastmediated mutagenesis system (Table 2). After cocultivation, 4 carcinogenic hydrocarbons [7,12-dimethylbenz(a)anthracene (DMBA), benzo(a)pyrene (BP), 3-methylcholanthrene (MCA) , and 7-methylbenz(a)anthracene (7-MBA)] induced ouabain-resistant mutants, whereas 5 noncarcinogenic hydrocarbons [benzo(e)pyrene, benz(a)anthracene, phenanthrene, pyrene, and chrysene] were not mutagenic.
The use of human cells and tissues for carcinogen activation in mammalian cell mutagenesis systems has been described by Harris and colleagues (26,27). Both human pulmonary alveolar macrophages and human bronchus tissue metabolized BP and its proximate metabolite, the 7,8-diol, into intermediates wh~ch converted V79 cells from ouabain sensitivity to ouabain resistance. Metabolities of BP were identified by high pressure liquid chromatography and the 54 ELIEZER HUBERMAN AND ROBERT LANGENBACH amount of BP binding to bronchial DNA was also measured in these studies.
Also, the choice of the medium in which the bacteria are treated may be of great importance, especially if the test agent requires active transport into the cells. This might not occur in buffer suspension. When testing agents in suspension culture, the protocol outlined below is followed: • A determination of the toxic level of the test agent in minimal essential medium is made. • After a determination of the toxic level, a pilot experiment is carried out using concentrations at either side of 50% toxicity level (total of five concentrations).
Application of Short-Term Bioassays in the Fractionation and Analysis of Complex Environmental Mixtures by Herbert S. Rosenkranz, Elena C. McCoy, Monica Anders, William T. Speck, David Bickers (auth.), Michael D. Waters, Stephen Nesnow, Joellen L. Huisingh, Shahbeg S. Sandhu, Larry Claxton (eds.)