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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