United States
Environmental Protection
Agency
Environmental Monitoring Systems
Laboratory
Las Vegas NV 89193-3478
Research and Development
EPA/600/S4-90/011 Aug. 1990
&EPA Project Summary
User's Guide-Micronucleus
Assay Data Management and
Analysis System
Raymond E. Tice and Al Pellom
This user's manual provides
guidance to researchers and the
regulatory community for interacting
with a data management and
statistical analysis system, desig-
nated as MN. MN is dedicated to the
in vivo micronucleus test, a routinely
used genetic toxicology assay for
chemical compounds which may be
of health concern. The objective in
developing this system has been to
promote consistency and inter-
comparability of assay test results
across laboratories, thus providing
researchers and government de-
cision makers with a means to assure
comparable analyses of test data.
The MN data management and
analysis system has been developed
in consultation with a panel of
biostatisticians and experts in the
field of cytogenetics. Software for
executing MN and three sets of test
data, contained on two 5.25 inch
floppy disks, accompany the user's
guide.
This Project Summary was
developed by EPA's Environmental
Monitoring Systems Laboratory, Las
Vegas, NV, to announce key findings
of the research project that is fully
documented in a separate report of
the same title (see Project Report
ordering information at back).
Introduction
MN is a data management and analysis
system designed for micronucleus data
collected using in vivo test systems. The
software consists of a set of routines for
1) entering, editing and storing
experimental data and descriptive
information; 2) generating statistics
appropriate to the micronucleus test; and
3) presenting the results of these
statistics through graphs and tables.
The user's guide consists of two parts.
The first describes the MN installation
program provided on the program disk.
The guide should be read carefully to
ensure that installation is performed
correctly. The second part describes how
to use MN. The description follows the
organization of the program, describing
the main menu first, followed by the
individual routines corresponding to each
of the menu entries.
Procedure
MN is intended to run on an IBM PC
compatible with hard disk under DOS.
The program files take up approximately
300K of storage on the hard disk, while
the sample test data require about 175K.
The installation procedure creates two
directories, C:\MN and C:\MN\HELP, and
copies executable and help files into
these directories. Copying the three test
data files into the C:'MN directory
simplifies recalling these data when
needed; otherwise, these files may be
recalled from the second floppy disk.
Operation
Each MN session maintains two
distinct data sets. The first is a multi-
page fixed form containing descriptive
information relating to the several aspects
of the experiment. The second is a
spreadsheet for the experimental data
(entered from current experiments or
from one of the test data sets), where the
rows represent observations and the
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columns represent fields. This form has a
number of fixed fields and a number of
optional fields that may be selected to
tailor a session to the nature of the
experiment. Help screens are available
within each menu selection.
The main menu (Figure 1) is a single
selection menu. When a selection is
made, the menu closes and the selected
operation begins. All but one of the
sixteen items of the main menu are
grouped into six classes: Setup, Data
entry, Disk I/O, Analysis, Utility, and
Miscellaneous.
Leave MN
This is a special item outside the six
functional categories. Selecting this item
will prompt for disposition of any unsaved
changes and return to DOS.
Setup
The Setup routines (Cell Types and
Optional Fields) determine which of the
nondefault fields will appear on the
spreadsheet. Cell Types allows the
selection of any combination of four cell
types for micronucleus analysis and one
of two configurations for toxicity analysis.
Optional Fields provides a way to define
up to fifteen additional data fields.
Data Entry
The Data Entry routines (Experiment
Description and Spreadsheet) are the
means by which data are entered into
MN sessions by keyboard. Experiment
Description consists of a six-page form
with which to enter descriptive
information pertaining to the experiment,
test article, solvent, positive control, test
system, and treatment. The spreadsheet
allows access to the actual data.
Disk I/O
Recall reads an MN file from hard or
floppy disk and makes it current. Save
writes the current MN file to disk. Import
and Export (not yet implemented) will
allow equivalent operations on ASCII files,
providing a means to move data between
MN and other applications.
Analysis
Statistics conducts specialized tests on
selected cell types and presents the
results via screen or printer. Graph plots
the means of selected response
variables.
Utility
List prints the data with the form of the
report subject to considerable user
control. Sort reorders the lines of the
spreadsheet (and optionally subsequent
listings) according to the values of any
combination of key fields, either
ascending or descending. Clear Session
deletes all data and removes all Setup
fields, in effect starting over.
Miscellaneous
Key Field Search produces a report
listing either 1) the values of selected
fields for all MN files in the indicated
search path, or 2) the names of all MN
files in the indicated search path whose
values for selected fields match key
values which have been entered. GLP
Log presents by screen or printer a
record of all changes ever made to non-
empty fields for the current file, who
made them and when.
Sample Data Sets
To assist the user in becoming familiar
with MN, three sample experiments
(MNTEST-1, -2, -3) have been included.
MNTEST-1 presents data on an
experiment in which male and female
mice were injected with a test chemical
on three consecutive days, bone marrow
samples were collected 24 hours after
the last injection and numbers of MN-
PCE (micronucleated polychromatic
erythrocytes) and the percentage of PCE
were determined by a single scorer.
MNTEST-2 presents data on an
experiment in which male and female
mice were treated once with a test
chemical, bone marrow samples were
collected at 24, 48, and 72 hours after
treatment and numbers of MN-PCE, MN-
NCE (micronucleated normo-chromatic
erythrocytes) and the percentage of PCE
were determined by two scorers.
MNTEST-3 is identical to MNTEST-2
except that the control data have been
restructured to only the 48-hour sample
time.
Software Availability
The Micronucleus Assay Data
Management and Analysis System
software can be obtained by sending two
formatted 5.25 inch 360KB disks to the
following address:
Dr. Charles H. Nauman
U.S. EPA, EMSL, MC-EAD
P.O. Box 93478
Las Vegas, NV 89193-3478
For further information the Micronucleus
Assay Data Management and Analysis
System source code and programmer
documentation, contact:
EPA Software Development Project
Integrated Laboratory Systems
P. 0. Box 13501
Research Triangle Park, NC 27709
I—Setup 1
Cell Types
Optional Fields
Data Entry -]
Experiment
Description
Spreadsheet
Leave MN
Recall
Save
Import
Export
[— Analysis —|
Statistics
Graph
r Utility
List
Clear
Session
Sort
r— Misc. —,
Key field
Search
GLP Log
Figure 1. Main Menu.
. S. GOVERNMENT PRINTING OFFICE: 1990/748-012/20086
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R. E. Tice and A. Pellom are with Integrated Laboratory Systems, Research
Triangle Park, NC 27709.
Charles H. Nauman is the EPA Protect Officer (see below).
The complete report, entitled "User's Guide-Micronucleus Assay Data
Management and Analysis System," (Order No. PB-90-212-598AS; Cost:
$17.00 subject to change) will be available only from:
National Technical Information Service
5285 Port Royal Road
Springfield, VA22161
Telephone: 703-487-4650
The EPA Project Officer can be contacted at:
Environmental Monitoring Systems Laboratory
U.S. Environmental Protection Agency
Las Vegas NV 89193-3478
United States Center for Environmental Research
Environmental Protection Information
Agency Cincinnati OH 45268
Official Business
Penalty for Private Use $300
EPA/600/S4-90/011
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