xvEPA
EPA-6DO/1-78-038
May 1978
NC J:.M
Development
Biomedical Data
Validation Through
an On-Line
Computer System
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RESEARCH REPORTING SERIES
Research reports of the Office of Research and Development, U.S. Environmental
Protection Agency, have been grouped into nine series. These nine broad cate-
gories were established to facilitate further development and application of en-
vironmental technology. Elimination of traditional grouping was consciously
planned to foster technology transfer and a maximum interface in related fields.
The nine series are:
1. Environmental Health Effects Research
2. Environmental Protection Technology
3. Ecological Research
4. Environmental Monitoring
5. Socioeconomic Environmental Studies
6. Scientific and Technical Assessment Reports (STAR)
7. Interagency Energy-Environment Research and Development
8. "Special" Reports
9 Miscellaneous Reports
This report has been assigned to the ENVIRONMENTAL HEALTH EFFECTS RE-
SEARCH series. This series describes projects and studies relating to the toler-
ances of man for unhealthful substances or conditions. This work is generally
assessed from a medical viewpoint, including physiological or psychological
studies. In addition to toxicology and other medical specialities, study areas in-
clude biomedical instrumentation and health research techniques utilizing ani-
mals — but always with intended application to human health measures.
This document is available to the public through the National Technical Informa-
tion Service, Springfield, Virginia 22161.
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EPA-600/1-78-038
May 1978
BIOMEDICAL DATA VALIDATION THROUGH
AN ON-LINE COMPUTER SYSTEM
by
Larry Claxton
Biochemistry Branch
Environmental Toxicology Division
Health Effects Research Laboratory
Research Triangle Park, N.C. 27711
U.S. ENVIRONMENTAL PROTECTION AGENCY
OFFICE OF RESEARCH AND DEVELOPMENT
HEALTH EFFECTS RESEARCH LABORATORY
RESEARCH TRIANGLE PARK, N.C. 27711
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DISCLAIMER
This report has been reviewed by the Health Effects Research
Laboratory, U.S. Environmental Protection Agency, and approved for
publication. Mention of trade names or commercial products does
not constitute endorsement or recommendation for use.
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FOREWORD
The many benefits of our modern, developing, industrial society are
accompanied by certain hazards. Careful assessment of the relative risk
of existing and new man-made environmental hazards is necessary for the
establishment of sound regulatory policy. These regulations serve to
enhance the quality of our environment in order to promote the public
health and welfare and the productive capacity of our Nation's population.
The Health Effects Research Laboratory, Research Triangle Park,
conducts a coordinated environmental health research program in toxicology,
epidemiology, and clinical studies using human volunteer subjects.
These studies address problems in air pollution, non-ionizing
radiation, environmental carcinogenesis and the toxicology of pesticides
as well as other chemical pollutants. The Laboratory participates in
the development and revision of air quality criteria documents on
pollutants for which national ambient air quality standards exist or
are proposed, provides the data for registration of new pesticides or
proposed suspension of those already in use, conducts research on
hazardous and toxic materials, and is primarily responsible for providing
the health basis for non-ionizing radiation standards. Direct support
to the regulatory function of the Agency is provided in the form of
expert testimony and preparation of affidavits as well as expert advice
to the Administrator to assure the adequacy of health care and surveillance
of persons having suffered imminent and substantial endangerment of
their health.
This paper presents how quality assurance controls were included
within the computer programming for a short term test -- the Salmonella
suspension assay for mutagenesis.
F. G. Hueter, Ph. D.
Acting Director,
Health Effects Research Laboratory
n
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INTRODUCTION
Within the biomedical disciplines there are a variety of testing pro-
cedures used routinely within many separate laboratories. Since health,
research and regulatory decisions are being based upon the results from
many laboratories, there is a basic need for assuring the quality of the
data. In the area of microbial mutagenesis, the use of Salmonella
typhimurium as an indicator organism for mutational events is employed
by many laboratories across the country. The various procedures available
are rapid, relatively simple, sensitive and are used in a variety of
laboratory situations including private industry, government and university
laboratories. Presently, a great deal of emphasis is placed upon these
types of tests as prescreens for substances that may be human mutagens and
potential carcinogens. Therefore, the use of a system involving Salmonella
typhimurium could provide an excellent pilot study for methods involved in
data validation. Data validation is used in this context to mean the
process by which generated data is filtered and accepted or rejected by
objective criteria. Likewise, computerization provides a potential
means for systematically applying a predetermined set of objective
criteria in a rapid non-biased manner. With the use of TSO (Time
Sharing Option), portions of the data validation can be conducted during
the performance of a biological test. This article will describe the
design of a pilot system for the on-line computer assistance of testing
protocols and data validation. The scientific protocols and initial
computerization have been completed and the system will be tested in a
laboratory situation in the near future by the National Institute of
Environmental Health Sciences.
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DESCRIPTION OF TEST:
From a variety of microbial mutation test systems, the suspension
test using a mammalian activation system was chosen because it is well
defined and is a quantitative test system.^ ' The more commonly used
Ames plate incorporation method is only semi quantitative. We also chose
to compare three strains of Salmonella typhimuriutn and a forward muta-
(?}
tion strain of K-12 E. £0_lj. ' In simple terms, the test involves the
combining of the bacterial strain with a compound and a mammalian activation
system into an Erlenmeyer flask which is incubated at 37°C for 30 minutes
to 2 hours. The bacteria are then separated and aliquots are plated on
minimal media for the detection of mutants and on supplemented media for
relative survival. Figure 1 provides a representation of the pilot test
presently used. Pilot tests are used to define more appropriate testing
conditions, and definitive tests provide data from which mutagenicity is
judged. For complete testing, the substance must be tested in several
strains of bacteria to monitor for a variety of different types of
genetic alteration.
SYSTEMS OVERVIEW
This program uses TSO and was written in COBOL with some additional
FORTRAN being integrated into the final program. All programming was
accomplished on an IBM System/370 at the Division of Computer Research
and Technology within the National Institutes of Health, Bethesda,
Maryland.
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For ease of programming, the task was divided into three individual
programs (Figure 2). Information, needed prior to testing of a parti-
cular substance, is stored with the use of Program 1. This program also
supplies a number for the blind coding of the compound. The second
program provides for the technician the proper form of the basic proto-
col, performs certain "within-experiment" calculations, accepts the
input of data from the tests, and evaluates the test by predetermined
objective criteria. The ability for the central laboratory to monitor
the accomplished work and recall any pertinent data is provided by
(3}
Program 3. A more precise description of the program is available.
Quality Control Through Interactive Computerization
One of the basic premises of quality control is that good data
yields good decisions. By monitoring the quality of data during an
experiment and providieng feedback to the technical personnel, both
personal bias and technical variation can be reduced. With an inter-
active computer network this can be done. This pilot project demon-
strates these capabilities in several ways. First, the compound to be
tested is coded and only essential information for the test is provided.
Secondly, certain other variables, e.g., concentrations of various
components, are predetermined for both the pilot tests and definitive
test. Within this testing system, two pilot tests are conducted to
determine levels of toxicology and potential mutagenicity. From this
data a narrower range of concentrations for the definitive tests are
calculated by predetermined rules so that there are a limited number of
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definitive concentrations used across all laboratories. Next, the
computer performs any needed calculations during the performing of a
test thus lessening the occurrence of potential computational errors.
Some of the calculations performed for this system are: (1) bacteria
per ml solution based on a standardized spectrophotometer curve, (2)
variance for the weights of animals used in microsomal S-9 preparation
(if outside normal limits, these will be rejected), (3) calculation of
liver weights and amounts of buffers to be used in microsome prepara-
tion, and (4) calculations for the dilution of samples. Final data
validation is also performed automatically upon the final data output.
The computer's ability for data storage and retrieval is very important
in this regard. For example, in this system, final results are recorded
as number of colonies per plate. This software program compares the
average number of colonies per plate for the controls to the past 100
accumulated controls to determine statistically if the controls are
within normal limits. After the statistical examination of the controls
the test is either accepted or rejected. If the test is a pilot then
the data is also used to determine the concentrations of test substance
to be used in further testing. All data is, however, recorded per-
manently. Rejected data is recorded so that an analysis of problems
encountered can be done. A flow diagram for the areas within the de-
cision processes is shown in Figure 3. The TSO is the component that
allows for immediate technician/program interaction, thus allowing for
a rapid and constant qualtiy control.
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This prototype system demonstrates that interactive computer pro-
grams can be used to effectively increase the quality control of rapid
jjn vitro tests. However, it is al~o apparent that the more simple
IB vitro microbial mutagenesis tests such as spot tests and simple plate
incorporation tests do not require such extensive computerization if well
documented and detailed protocols are available. Since most J_n vivo
mammalian systems have extended experimental time periods, the time sharing
option would be of little benefit due to cost factors and experimental
design. However, even with the more simple i_n vitro tests and mammalian
cell culture tests, this system can serve as a model for data storage
and test evaluation for the purpose of quality control.
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REFERENCES
1. Frantz, C. N. and Mailing, H. U. 1975. The Quantitative Microsomal
Mutagenesis Assay Method. Mutation Research 31:365-380.
2. Mohn, Georges, Ellenberger, J. and McGregor, D. 1974. Development
of Mutagenicity Tests Using Escherichia coli K-12 As Indicator Organism.
Mutation Research 25:187-196.
3. Claxton, Larry and Baxter, Richard. 1978. The Computer Assisted
Bacterial Test for Mutagenesis. Mutation Research (In Press).
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TECHNICAL REPORT DATA
iPlease read lusln tenons on the reverse before completing)
1 REPORT NO 2
EPA-600/1 -78-038
4. TITLE AND SUBTITLE
Biomedical Data Validation Through an On-line
Computer System
7 AUTHOR(S)
Larry Claxton
9. PERFORMING ORGANIZATION NAME AND ADDRESS
Environmental Toxicology Division
Health Effects Research Laboratory
Research Triangle Park, N.C. 27711
12. SPONSORING AGENCY NAME AND ADDRESS
Health Effects Research Laboratory RTP , N(
Office of Research and Development
U.S. Environmental Protection Agency
"RciQpprrh Trnanalp Park., N.C. 27711
15, SUPPLEMENTARY NOTES
3. RECIPIENT'S ACCESSION-NO.
5. REPORT DATE
May 1978
6. PERFORMING ORGANIZATION
8. PERFORMING ORGANIZATION
CODE
REPORT NO.
10. PROGRAM ELEMENT NO.
1AA6.01
11. CONTRACT/GRANT NO.
13. TYPE OF REPORT AND PERIOD COVERED
-1
14. SPONSORING AGENCY CODE
EPA 600/11
16. ABSTRACT
Since health and regulatory decisions are being based upon the results
many short, term tests conducted in many laboratories, a computerized system
for an assurance of quality control would be valuable. This paper presents
how quality assurance controls were included within the computer programming
for a short term test — the Salmonella suspension assay for irutagenesis.
of
17. KEY WORDS AND DOCUMENT ANALYSIS
a. DESCRIPTORS b-IDENTIFI
data processing data v
data reduction
computer systems programs
biomedical measurement
13. DISTRIBUTION STATEMENT 19. SECURI
UNCLA
KKLKAbE TO PUBLIC 20 SECURI
UNCLA
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alidation 06 B
leld/Group
TY CLASS (TMsReport) 21. NO. OF PAGF1
SSIFIED 13
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EPA Form 2220-1 (9-73)
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