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Genotoxic exposure assessment by simplified DNA analysis.

Authors
Prichard-HM
Source
School of Public Health, University of Texas, Houston, Texas 1988 Nov; :1-5
Link
NIOSHTIC No.
00187455
Abstract
A test which was both sensitive and practical was sought to study human exposures to genotoxic agents. The test was based on the changes apparent in the 340 base pair (bp) alphoid sequence subsequently brought about by Eco-R1 restriction of DNA obtained from peripheral leukocytes. Following in-vitro challenge by ionizing radiation or by various chemical agents, the DNA was extracted from human leukocytes. Breakage was induced at the damaged sites of the 340bp alphoid sequence by subjecting it to Eco- R1 restriction and secondary treatment. The distribution of a labeled 340bp probe following electrophoresis and hybridization was used to determine the degree of breakage. When appropriate it may be possible to use other means to quantify the breakage. Efforts were successful in keeping the quantity of blood needed for the analysis to a small amount. Electrophoresis, followed by cerenkov counting, was effective in detecting the action of in-vitro exposures of dimethyl-sulfoxide (67685) on leukocytes. DNA samples extracted from cells exposed to four treatment levels were restricted, radiolabeled, made alkaline, and subjected to gel electrophoresis. Tracks were divided into three regions by excision of the 340bp region and the activity of each was assayed by cerenkov counting. The fraction of the labeled genome heavier than 340bp fell monotonically from 93 to 3 percent as the dimethyl-sulfoxide concentration was raised. The carcinogen N-methyl-N'-nitro-N- nitrosoguanidine (70257) produced similar, but less pronounced, results.
Keywords
NIOSH-Grant; Reproductive-system-disorders; Nucleic-acids; DNA-damage; Genotoxic-effects; Analytical-methods; Chemical-analysis
Contact
Environmental Sciences University of Texas P O Box 20186 Houston, Tex 77225
CAS No.
67-68-5; 70-25-7
Publication Date
19881123
Document Type
Final Grant Report
Funding Amount
94938
Funding Type
Grant
Fiscal Year
1989
NTIS Accession No.
PB89-201008
NTIS Price
A01
Identifying No.
Grant-Number-K01-OH-00035
NIOSH Division
OEP
Priority Area
Reproductive-system-disorders
Source Name
School of Public Health, University of Texas, Houston, Texas
State
TX
Performing Organization
University of Texas Hlth Sci Ctr Houston, Houston, Texas
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