EPA/AA/CTAB/PA/81-10
                  Brief Synopsis of EPA
           Office of Research and Development
                         and the
       Health Effects Institute Mobile Source Work
                        May, 1981
Control Technology Assessment and Characterization Branch
          Emission Control Technology Division
      Office of Mobile Source Air Pollution Control
           Office of Air, Noise  and Radiation
          U.S. Environmental Protection Agency
                   2565 Plymouth Road
               Ann Arbor, Michigan  48105

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Summary

The EPA Office of  Research  and Development has many  important mobile  source
projects  in  progress as  discussed  in this  paper while  the Health  Effects
Institute will begin work shortly.   Some  of the  more  important  items  that
can be noted at this point are given below.

1.  ORD-ESRL has developed  a  preliminary method to collect  gas  phase  hydro-
    carbon samples  from gasoline and Diesel  vehicles for bioassay  testing.
    This method involves use of XAD-2 cartridge traps and  is presently being
    written up.  After  the method  is available  later  this  year, work  can
    start to make an assessment of the mutagenicity of the gas phase  samples
    relative to the particulates.

2.  ORD-ESRL  has  done  a  number  of  tests   to  determine   if  N0£   in  the
    exhaust gas  can  cause  artifact  mutagen  formation  on  the  particulate
    filter and  thus an artificially  high Ames  test  response.   Preliminary
    results indicate that no  artifact will be  formed provided that the  N0£
    levels in the diluted exhaust are below 5 ppm which  is  usually the  case.

3.  Significant work is being  done to  identify  the compounds responsible  for
    the   mutagenic   activity   in   Diesel   exhaust.    Numerous   oxygenated
    polynuclear  aromatic  hydrocarbons   and   nitroaromatics   have    been
    identified as being responsible for much  of the mutagenic activity.

4.  Additional light  duty  Diesel  and gasoline  vehicles  have  been  tested.
    Particulate emissions and  Ames  test  response  have been  measured.   Also,
    several heavy duty  Diesel  engines have been  tested.   Much work will  be
    done  on various  synthetic  fuels  derived  from  coal  and  oil-shale  in  the
    future.

5.  ORD-EMSL  has  some  short  term  monitoring  work  underway   to  measure
    personal exposure to CO and correlate it  to the readings  taken at  fixed
    site NAAQS monitors.  This work involves  contractors measuring CO  levels
    along commuting  routes  and other  places  (e.g. central business  district
    and outlying areas).

6.  A longer term  CO monitoring project involving a  large number of  people
    wearing CO monitors should be  started soon.   Their  CO exposure will  be
    correlated to  their personal  activity and to readings  obtained  at  the
    fixed site NAAQS monitors.

7.  Short term bioassay results have been  obtained on a number of Diesel  and
    other  samples   including   coke  oven,  cigarette   smoke  condensate,   and
    roofing tar emissions.

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                                   -2-
7.  Short term bioassay results have been  obtained  on  a number of Diesel and
    other  samples  including  coke oven,  cigarette   smoke  condensate,  and
    roofing tar emissions.
8.  Skin  tumorigenesis  initiation data  are  available  on  Diesel  particulate
    samples  and  indicate  these   samples  are  less potent  than  coke  oven
    samples.  Intraperitoneal  injection  results should  be  available shortly
    but the  intratracheal instillation  results will not be available  until
    1983.

9.  Results so  far on whole  animal  inhalation work with Diesels have been
    generally negative with the  exception of some  sister  chromatid  exchange
    work with hamsters.   The sister  chromatid exchange  work showed  a linear
    relationship between  Diesel  exposure  and  the  sister  chromatid  exchange
    response.

10. -ORD-HERL  has   started  a  clinical  study  to establish  conclusively  the
    relationship between CO exposure and the decrease  in time  until  onset of
    chest  pain  for  people  with  angina  pectoris  during  exercise.   Also,
    animal  work  is  starting  to  determine  the  effect  of  CO  exposure  on
    fetuses.  Results for  the clinical and animal  studies will be available
    in 1982.

11. ORD-HERL  is doing  work to develop a tier bioassay  test  to be  used  for
    mutagenic  and  carcinogenic   substances  for  OMSAPC  regulatory  efforts
    under  Section  211  (fuel  and fuel  additives)  and  Section  202  (a)(4)
    (assuring  motor  vehicle  systems   being  certified  do   not   cause  or
    contribute to  an unreasonable risk  to public  health or welfare) of  the
    Clean Air Act.  The tier  test protocols  are expected to be available in
    late 1982.

12. A revised risk assessment estimating  the  potential  carcinogenic  impact
    of  Diesel exhaust  emissions  is  being prepared  by  ORD   and  should  be
    available shortly.   Meanwhile, ORD has concluded that  the  London Transit
    Worker  study   is not   adequately  sensitive to  show  there would  be  no
    excess  cancer  deaths   resulting  from  the  projected  exposure  to  Diesel
    exhaust.

13. The newly organized Health Effects  Institute  is  an  independent  organi-
    zation designed to conduct health  research on motor  vehicle  pollutants.
    The Institute  is  currently planning the  programs  they plan to  conduct;
    EPA  and  the   motor   vehicle   industry will  be  submitting  a  list  of
    suggested program areas to the Institute.

Please call me if you have any questions or comments on this report.

Addressees:   Flo Ryer       Ron Bradow
              Gerry Rausa     Joellen Lewtas
              Deran Pashan   Judy Grahm
              Lance Wallace  Don Horstman
              Gerry Akland   Bill Pepelko

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                                      -3-


8.  Skin tumorigenesis  initiation  data are  available  on Diesel  particulate
    samples  and  indicate  these  samples  are  less  potent  than  coke  oven
    samples.  Intraperitoneal  injection  results should be available  shortly
    but the  intratracheal  instillation results will not  be available  until
    1983.

9.  Results so  far  on whole  animal  inhalation work with  Diesels have  been
    generally negative with  the  exception of some sister chromatid  exchange
    work with hamsters.  The sister  chromatid exchange work showed  a  linear
    relationship between Diesel  exposure and  the  sister chromatid  exchange
    response.

10. ORD-HERL has  started  a   clinical  study  to establish  conclusively  the
    relationship between CO exposure and  the decrease  in time  until  onset  of
    chest  pain  for  people with  angina pectoris  (a  heart  condition)  during
    exercise.   Also,  animal  work  is  starting to  determine  the effect of  CO
    exposure on  fetuses.   Results  for the  clinical  and animal studies  will
    be available in 1982.

11. ORD-HERL is  doing work to develop a  tier bioassay test  to be used  for
    mutagenic  and  carcinogenic  substances  for  OMSAPC  regulatory  efforts
    under   Section  211  (fuel  and  fuel  additives)  and Section  202  (a)(4)
    (assuring  motor  vehicle  systems  being  certified  do   not   cause   or
    contribute to an  unreasonable  risk to public  health  or  welfare) of  the
    Clean  Air Act.  The tier  test  protocols  are expected to be available  in
    late 1982.

12. A revised risk  assessment estimating  the  potential carcinogenic  impact
    of  Diesel  exhaust  emissions  is  being  prepared  by ORD   and  should  be
    available shortly.  Meanwhile,  ORD has concluded that the  London  Transit
    Worker study  (a Diesel epidemiology   study)  is not adequately  sensitive
    to  show there  would  be  no  excess   cancer deaths  resulting  from  the
    projected exposure to Diesel  exhaust.

13. The newly organized  Health Effects Institute  is  an independent  organi-
    zation designed to conduct health research on motor vehicle  pollutants.
    The Institute is  currently planning   the  programs  they  plan to  conduct;
    EPA and  the motor  vehicle  industry  will  be  submitting  a  list   of
    suggested program areas to the  Institute.

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                                      -4-
I.  Introduction

For a number of  years,  the  EPA Office of Research and  Development  (ORD)  has
conducted a number of mobile  source  research  programs.   The initial programs
were  characterization  ones  done  by  the  Environmental  Sciences  Research
Laboratory (ESRL)  which were  followed in  about 1975  by mobile  source  pro-
grams in both the Health  Effects Research Laboratory  (HERL)  and the Environ-
mental Monitoring and Support Laboratory  (EMSL).

In  1978,  EPA started  the Mobile  Source  Research Committee  (MSRC) to  help
assure that ORD  mobile  source  work is  responsive to  the needs  of the  Office
of Mobile Source Air  Pollution Control (OMSAPC).  Since that time,  a  number
of problems  have surfaced.   OMSAPC  feels  that  ORD will sometimes  implement
work  ORD  feels  is  needed  rather  than work  specifically  requested by  the
program  office.    ORD  will   frequently  do   some   projects   that   OMSAPC
disapproves  since  OMSAPC  feels these  projects do  not meet   OMSAPC  needs.
Also, there  have been  problems  in transmitting results of ORD  research  to
OMSAPC.   On the  other hand, ORD frequently feels that  some  work requested by
OMSAPC  is  either  not  scientifically valid  or  too  short term  and  of  a
service-type function  (e.g.  run 300 samples  for Ames  test  activity)  rather
than  longer  term research.   Even with these problems,  there  is considerable
ORD mobile source  work that is useful to OMSAPC.  Also,  ORD and  OMSAPC  are
trying to resolve the problems experienced to date.

The  purpose  of  this  report  is not  to  discuss  any  problem  areas  between
OMSAPC and ORD but instead  to  cover  the ORD mobile source work  itself.   This
report will  summarize  the  work in  progress by  ORD  and  give  some of  the
pertinent  results.    This  report  will  also  briefly  mention  future  work
planned by ORD.

The results and work discussed  in  this paper are that  available to OMSAPC as
of April 10,  1981.

II. Chemical Characterization Work

The chemical characterization work  for  unregulated  mobile  source  emissions
is done by ESRL.  This  program involves developing methods  to measure  unreg-
ulated emissions and conducting extensive Diesel characterization work.
A.  Development of Method to Collect
    Gas-phase Hydrocarbons in Diesel Exhaust

One of the highest priorities expressed by OMSAPC was  to  develop  a method to
collect  artifact-free  samples  of  gas  phase  hydrocarbons  in motor  vehicle
exhaust  for  bioassay testing.   ESRL has evaluated  both  a  filter cartridge
and a condensate method.

Preliminary  tests have  been  done  with  a  method  involving filtering  the
exhaust particulates and then condensing  components  in the  gas  stream.   The

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                                      -5-
condensate appears  to  have low Ames  test  activity.   If the  filter  upstream
of  the  condenser is removed,  the  condensate will  contain some  Diesel  par-
ticulate and has somewhat higher Ames test activity.


The  filter  cartridge method  involves passing  a  gas stream  sample after  a
conventional particulate  filter  through  a cartridge or bed of  treated XAD-2
resin.  After  the  gas  stream  is passed  through the XAD-2, the hydrocarbons
absorbed onto  the  XAD-2  are  removed  from  the resin  by  methylene  chloride
extraction.  The  lower molecular weight hydrocarbons  (e.g.  below C-10)  are
sufficiently volatile  that they  are  probably  lost  during  the  extraction.
However, hydrocarbons above C-10 are  retained and can then be  subject  to the
Ames  test.   Since  the  conventional particulate filter will generally  retain
hydrocarbon  compounds   above  C-15,  the  XAD-2  traps  could   provide  a  good
method to collect hydrocarbons in the C-10 to C-15 range.

A very preliminary  result  of  Ames  testing on the gas phase hydrocarbon  col-
lected by this method  for  a VW Diesel Rabbit shows that the  activity  may be
low compared to that of the particulate.

The methods  and  results mentioned  above  are very  preliminary  and have  not
yet been published  by  ORD.  However,  OMSAPC  has  requested that  ORD  publish
these results with a full description of  the method as soon as possible.

In  the  past,  ORD has  expended great  effort  in identifying  the  hydrocarbon
compounds  in  the gas  phase  of Diesel  and  gasoline  vehicle  exhaust.   This
work has been described by ORD  in  several publications and was  recently  sum-
marized in an OMSAPC report. ^


B.  Potential Effect of N02 in
    Mutagen Artifact Generation

EPA has  been concerned that high  levels  of N02  in  the  dilution tunnel  may
result  in  a reaction  of  N0£ with  some  of  the hydrocarbons  present  either
in  the  gas  stream  or  on the particulate  filter.   The  reaction  products
including various nitroaromatic compounds  could result  in an increased  Ames
test  response.   Since  these   reaction products  from the hydrocarbons  and
M>2 would  not  be expected to  form in the  atmosphere  where  these  compounds
would be greatly diluted,  the Ames test response would be artifically  high.
ORD has  run  some  experiments where  additional  N0£ has been  introduced  into
the dilution tunnel.   These experiments  show  that artifact  is created  in
Diesel  particulates  if the  N02  levels  are  above  5 ppm.   ORD  plans  to
publish these results shortly.

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                                      -6-
A  concern has  also  been  raised  that  lower  levels of  N(>2 may  result  in
artifact  formation  although  this artifact formation would probably  be  lower
than that which  might be experienced at  higher  N(>2 levels.   OMSAPC  has  run
some experiments  with a single  cylinder  Diesel  engine with  artificial  com-
bustion  air   containing  no  nitrogen and a  nitrogen  free   fuel.   In  this
experiment,  no  NOx would  be formed  in  the exhaust  gas.   By comparing  the
results  of  these  tests  to  tests with  conventional  air  (80%  N2,  20%  0£)
and regular Diesel  fuel  (which  contains traces of  nitrogen), one  can deter-
mine  if  there  were  artifact  formation  due  to  the  NC^.    The  results  of
these tests have  not  yet bee: published.  Also, investigators at  Penn  State
University and  John  Hopkins  University  plan  to  repeat these  tests in  the
coming year.

C.  Study of Filter Efficiency

ORD has  just  completed a study^ examining various  filters  that can  be  used
for Diesel particulate collection.   The  filters  investigated  were  Millipore,
Fluoropore  FA,   Dexiglas,  Gelman  A-E,   Ghia  Zefluor   lu,  Pallflex  T60A20,
Pallflex TX40HI, and Pallflex 400HO.

A common back-up  filter  was  used to  measure any break-through from  the  test
filter.  It was  found that all  of the filters collected greater than 95% of
the  particulate  on  the  primary  filter  as  determined  by  the  standard  EPA
method^.   However,   it  was  noted  that  small  differences  in  collection
efficiencies  occurred in the cold start  bag  of  the Federal Test  Procedure
rather than the portions when the vehicle engines were warmed-up.

Even though all of  these fiters  may  be  acceptable  for determining  total  par-
ticulate mass, it  is  not known  if an interaction of the filter and  the  par-
ticulate  such  as can  occur  with glass  fiber  filters  would  affect  the  Ames
test results.

D.  Identification of Types of Compounds
    Responsible for Ames Test Activity in
    Diesel Particulates.

ORD has  spent effort  in  separating the methylene chloride extract of Diesel
particulates by  high  pressure  liquid chromatography.   Three  major  fractions
have been obtained:   non-polar,  moderately polar  (transition),  and  highly
polar.    Some of  this  work  has  been done  in  a  cooperative  scientific project
with Ford Motor Company.

The  non-polar peak  consists of  aliphatic  and  polynuclear   aromatic  hydro-
carbons  (many  of them alkyl substituted) which  are not  responsible for  a
major portion of the Ames test activity.

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                                       -7--
Th e  transition  and  polar  fractions  account  for  most  of  the  Ames  test
activity.  About  50% by weight  of the material  in the  transition  fraction
consists of  various oxygenated polynuclear aromatic  hydrocarbons  (including
hydroxy,   ketone,   carboxaldehyde,  quinone,   acid  anhydride,   and   nitro
compounds).   Three  specific  nitro  compounds  including  1-nitropyrene  have
been  identified.    A  total  of  about  80  other   compounds  have  also  been
identified.   The   remainder  of  the  transition fraction  is  thought  to  be
contaminants  introduced  in  the analysis procedure.  The  polar  fraction  con-
sists of  carboxylic acid polynuclear  aromatic  hydrocarbons  which also  have
significant Ames test activity.^

It appears that a  great  number of  compounds may be  responsible  for  the  Ames
test activity.

E.  Particulate Emission Factors From Light and Heavy Duty Vehicles

OMSAPC  expressed  a need for additional  particulate emission data  from  both
gasoline and Diesel vehicles.

Tests were  run in-house at ESRL  on a  1980  VW  Diesel Rabbit  and  Oldsmobile
Diesel.   Some tests were done  with  the  VW under malfunction  conditions.   The
table below summarizes some of the most pertinent  results.

                                    Table  1
                    ESRL Tests on VW and Oldsmobile Diesel

Vehicle       HC        CO       NOx       Particulate    Soluble Organics
              g/mile    g/mile   g/mile    g/mile                 %

VW normal     0.51      1.31     0.99      0.43                  27
VW - a        0.68      1.69     1.23      0.54
VW - b        0.74      1.70     1.28      0.76
VW - c        0.51      1.31     0.99      0.43
Oldsmobile    0.25      1.24     1.54      0.54                  15

         a    timing retarded/bad injector
         b    bad injector
         c    new injector

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                                      -8-
These test  results show that  a bad injector  increases the  emissions;  sim-
ilarly specific Ames test  activity  increased by  over 50% on  TA98 due  to  the
faulty  injector.   Tests  of  both  vehicles  over  differenmt  driving  cycles
showed that Ames test specific  activity was  affected slightly by the driving
cycle:  the  cycles examined were the  FTP, HFET,  New  York  City cycle,  and
Congested Freeway Driving Schedule^.

Additional  studies"  were  done  for  EPA  by the New  York State Department  of
Environmental  Conservation  on  19  in-use  Diesels  at  different  mileages.
These tests  showed that various driving  cycles> from  the  limited  number  of
tests run, do not affect Ames  test  specific  activity other  than  emissions  at
idle  are   somewhat  less   active.   These   tests   show  in-use   particulate
emissions appear to be in the  same  range  as  obtained from  tests  of  prototype
vehicles.   However,  the  results to date  may be too  limited  to  make  this  a
firm conclusion in part since little data at high mileage are available.

ESRL  has  also conducted  a contract study  at  Southwest Research  Institute?
to measure  unregulated  emissions from  four  heavy  duty  engines  (Caterpillar
3208, Mack  ENDT 676, Cummins  290,  and  Detroit  Diesel 8V-71)  over   the newly
developed EPA  heavy-duty-engine transient cycle.  A summary of  some  of  the
emissions results is given below.

                                    Table  2
                  Emissions Results From Heavy-Duty Engines*

Engine                  HC       CO       NOx       Particulate
                        g/mile   g/mile   g/mile    g/mile

1979 Caterpillar 3208   1.98     5.6      18.6       0.92
1979 Mack ENDT 676      1.61     9.6      29.1       1.95
1979 Cummins 290        2.37    11.2      25.4       1.64
1977 DD 8V-71           2.25    64.9      39.1       2.74
         *results obtained over 1983 transient  cycle
It was  found that  the  Los Angeles  Freeway segment of  the transient  cycle
contibuted most  (40-50%)  of the particulates  collected.   It was  found that
sulfur compounds  constituted  about  10%  of the  particulate mass  presumably
due to sulfate.   Also,  HCN and ^0  emissions  were  measured and found  to  be
below  80 mg/mile.   Emissions  were  measured  with  different  fuels;  it  was
found that  HC was higher  with  the minimum quality  DF-2  with  the  4-stroke
engines and benzo(a)pyrene (BaP) emissions also increased  with this fuel.

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F.  Particulate Emissions from In-Use Gasoline Vehicles

ESRL  started an  in-house  program to measure  particulate emissions  from 20
in-use  gasoline  vehicles.   Of these cars, 16  are  catalyst-equipped  and thus
burn unleaded fuel.  Ames tests will be  run  on the samples collected.  Tests
have so  far  been  run on 13  of the 20 cars but no  results  are  available yet.
A final report on this  is due later this year.

G.  Synthetic Fuels Work

ESRL has obtained  some  shale-oil  Diesel  fuel samples and  plans  to  test some
vehicles  in-house with them.  Both  gaseous  emissions  including  gas-phase
hydrocarbons and  particulates will  be  analyzed.   Some limited work  was done
with  a  relatively high quality  oil shale  derived  Diesel fuel   showing  the
Ames  test  reactivity   of  the particulates  is  roughly   equivalent   to  that
obtained with  conventional  fuel.8  Future  ESRL  plans   call  for  obtaining
H-coal,  SRC-II  and donor solvent  fuels.   This work complements  work OMSAPC
has  underway  on  coal  and  oil-shale  fuels.   Also,   the  ORD  Industrial
Engineering Research Laboratory (IERL)  plans to start a program here.

III. Carbon Monoxide and Other Monitoring Work

EMSL has a number  of monitoring projects underway for CO  and  also  does some
fuels  and  quality assurance  work.   Some of  these  are  short term  projects
while others are longer term.

A.  Carbon Monoxide Work

In  the  past several years,  several  studies  9,10,11,12,13,14,15 have  been
done  indicating  that  CO levels  that  people  are  actually exposed  to  are
greater than predicted  on the basis of the CO  NAAQS  fixed-site monitors.   In
particular, work^  has  shown that  the  fixed-site monitors underestimate  CO
exposure for 1  hour periods  for  Boston  commuters by about  a factor  of  2.
OMSAPC  has  placed  very high  priority   on  ORD work to  define  what  actual
exposure is to  CO.  While it  is recognized that the  EPA  NAAQS  network for CO
is good  for determining CO  trends  and  levels  at fixed sites,  the  fixed site
network is  not  as accurate as needed to determine actual  exposure.

Another  pollutant  EPA  needs  actual exposure  data on is  Diesel  particulates
so that  more accurate  carcinogenic risk  assessments  can be made.  While  it
is  not  currently possible   to  determine  actual  exposure  to  Diesel  par-
ticulates  by  having  people  carry  Diesel   particulate  monitors,   CO  can
possibly be used  as  a  surrogate  for Diesel particulate  exposure.   Thus,  the
CO personal  exposure work  can possibly be used  to refine the Diesel cancer
risk assessment.

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                                        -10-
OMSAPC has  requested that  ORD implement CO  exposure projects  in two  time
frames; a  short  term project  can provide OMSAPC  with preliminary data  for
OMSAPC1 s more  immediate needs while  a  longer term project  can look  at  the
problem in more detail and meet longer term needs.

The short term CO monitoring project  is  being  run  in  three cities,  Stamford,
Connecticut,  Denver,  and  Phoenix.   ORD  contractors  are  carrying  a  CO
personal monitor  while  driving over  typical  commuting routes  determined  by
local  transportation data.   The monitors  are  carried   along  these  routes
during morning and  evening rush hours.   An  average CO exposure value (e.g.
10 ppm 1 hour average)  is  determined  for certain time periods.   Then, during
the rest  of the day, CO  values are  obtained  by walking around the  central
business  district   and  driving  in  outlying  suburban areas.   Readings  are
taken in various public buildings and stores.

These  data  are  representative of what  CO levels  people  are exposed  to  and
will  be  compared  with  values  recorded  at  the NAAQS monitors.   The  data
should be  collected by May 1981 and will  be  analyzed   shortly  thereafter.
Some results should be available later this  summer  from this analysis.

Other  EMSL  short  term work being  done on CO  monitoring  include a  contract
with  Sciences  Application  Inc.  who  has outfitted nine  people  in   the  Los
Angeles area with  CO monitors  and  recorded  their  CO  exposure  as they  went
about  their normal  activities.   Data  from  this  study  should  be  available
later this year.

EMSL  is also planning  a longer term  CO  monitoring study which will  include
an assessment of people's  activities  as  well  as their CO  exposure.   A diary
type  log  will  be kept  by  the people carrying  the monitors to record their
activities.  Thus,  a relationship can be developed between personal  activity
and CO exposure.   It  is  tentatively expected that  about  100 people  will
carry the monitors  for  certain time periods.   This project  will give OMSAPC
a more definitive assessment of  actual  CO exposure as compared to that  pre-
dicted from the  fixed site NAAQS  monitors.   This  work  is starting  in  FY81
and will continue into FY82.

Some  limited FY81  work is  underway to  improve CO personal monitors  and  to
incorporate  integrators in  them  so that  both instantaneous  CO  and  time
integrated CO measurements can be taken.

B.  Fuel Registration and Analysis

EMSL  has  the  responsibility for  operation  of the   fuel registration  program
required by Section 211(b) of  the  Clean Air  Act  Amendments.  This  program
involves  registration  of all   fuel  and   fuel  additives  listing  their
components, concentration  levels,  and purpose of using the  additives.  Much
of  the information  (especially  on  fuel additive  composition)  can   be  held
"confidential" by the provisions of these regulations.  EMSL plans  to issue
a report on the fuels and  fuel additives registered in late 1981.

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                                      -11-

Other  provisions  of  Section  211  require  EPA  to  promulgate  regulations
requiring needed health or  emission-control-device  testing  of  fuels  and fuel
additives.   Initially,  ORD  was developing  these  regulations  but   in  early
1980  the responsibility  for  proposing  these regulations was  shifted  to
OMSAPC.  OMSAPC is still counting on ORD to  develop  a  tier  bioassay  test for
mutagenic compounds;  this work  is  discussed  later  in Section  IVC.   OMSAPC
will  use these  tier bioassay  tests  as one element  in  the  fuel   and  fuel
additive protocols.

EMSL  also provided analysis  support  to the Office  of Enforcement  on  trace
contaminants in unleaded  fuel such  as  lead, manganese  and  phosphorus.   EMSL
also  analyzed  some   fuel   samples   for   OMSAPC,   performed  benzo(a)pyrene
analyses on  Diesel  particulate samples, and ran sample extractions  prior to
Ames  test analysis.   However, EMSL  management decided  the  priority  for this
work was low and recently discontinued these analyses.

IV  Health Work

The ORD-HERL Laboratories in  Cincinnati and Research  Triangle  Park provide
OMSAPC with  research  support in the mobile  source health area.   The current
work  is on Diesels and development of a tier biossay test.

A.  Diesel Work

The largest  amount  of HERL FY81 mobile source resources  are being  spent on
Diesel  health work.   The  work  includes  short  term  bioassay tests,  skin
tumorigenesis,  intratracheal  instillation,  and  whole  animal  inhalation.
This  work is  being  used to  determine a  carcinogenic  potency for  Diesel
exhaust  particulate  to  use  in  a  carcinogenic  risk  assessment.   The  end
product  of  all of  the  Diesel health work will be  a cancer risk  assessment
predicting the carcinogenic  risk  associated  with exposure to light  and heavy
duty  Diesel  exhaust.   The health  work on  Diesels is discussed  below but the
Diesel risk  assessment is covered in Section V as a separate entity.

1.  Bioassay Tests

    For  several  years  HERL-RTP has  been  running  a variety  of short  term
    bioassay  tests  including  the  Ames   test,  sister  chromatid  exchange,
    L-5178Y  mouse  lymphoma, and Balb  tests.  Two  short  term  carcinogenesis
    tests (viral enhancement  and Balb) have  also been run.

    These tests  have been  run on a variety of  light  and  heavy  duty  Diesel
    particulate  extracts.   The light  duty  Diesels  tested  include  an  Olds-
    mobile,   VW Rabbit,  and  Nissan.   The Nissan Diesel  had  a poorly  designed
    injector  (which  has   been  redesigned  on newer   Nissan  Diesels)  that
    resulted  in  considerable  "after  injection"   (i.e.   late   injection)  of
    Diesel  fuel.   EPA understands  that  Ames  test  results   with  the  new
    injection   system  are    considerably   lower.    The  samples  from  the
    Caterpillar heavy duty  Diesel were stored for an  extended period which
    may  have resulted in sample  deterioration.   OMSAPC has requested  that

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                                   -12-


ORD  run  new heavy duty  Diesel  samples.  Bioassay  tests have  also  been
run  on  particulate extract  from  a limited number of  gasoline  vehicles.
The  gasoline  car  tested was a  1977  Mustang  catalyst  equipped  vehicle
that apparently  was  running  somewhat  richer than manufacturers'  speci-
fications.

To  date,  very  few if  any bioassay tests  have  been  run  on  gas  phase
samples from either gasoline  or  Diesel vehicles mainly due to  lack  of a
method to  collect  samples for bioassay tests.  However,  as mentioned in
Section II  A,  work is proceeding by ESRL  to develop  such a method and
one is expected shortly.

The  bioassay  test response  of  the  motor vehicle  samples  have  been
compared  to other samples.   The  other samples  collected are  from  coke
ovens,  roofing  tar,  and  cigarette  smoke  condensate.   The  coke  oven
samples collected by ORD were taken from a  coke  oven owned  by U.S.  Steel
in  Gadsen,  Alabama.   The samples were  collected  in  a location  that was
upwind of  the  coke ovens a large  fraction  of  the time.   However, it is
thought that much  of  the sample mass  was  collected  when the coke  ovens
were discharging and presumably the wind was  in  the  right direction  even
though this may have  occurred  only a small fraction of the  time.   In
other words,  OMSAPC  is  not  currently sure how  much  of the  coke  oven
samples  are  actual   coke  oven  emissions.^   OMSAPC  hopes  that  this
point is clarified in the  ORD report to be issued on  this work.   ORD is
currently obtaining additional  coke  oven  samples for  bioassay  and  other
tests.  The additional  samples  are being  obtained in  the coke  oven  main
itself and  may  give  different results  from the earlier  samples.  It is
not known yet how  the new  samples  would be  representative of  actual  coke
oven  emissions  to  which  people  are  exposed.   The  cigarette  smoke
condensate  samples were obtained  by   a  standard   laboratory  apparatus
designed to duplicate cigarette smoking.  The roofing  tar emissions  were
obtained by collecting  a particulate  sample above a hot pot of  roofing
tar.

The reason  that  the coke oven,  cigarette  smoke,  and roofing  tar  samples
were  investigated  is  that  extensive  epidemiology data  exist  for  these
substances  while  almost no  good  epidemiology  data  exist  for  Diesel
emissions.

The results from the bioassay tests have been calculated as  the response
per  unit  mass of  particulate extract.  The  responses were  rated  on  a
relative basis with the  Nissan arbitrarily  assigned  a  ranking of  100.  A
known carcinogen  (benzo(a)pyrene)  was  used for comparison.   The  results
from  this  work  are  summarized in Table  3  below and are  discussed  in
detail in  several  references.^7,18,19,20   They were also covered in an
ORD   sponsored   workshop    held   February    1981    on    Diesel   risk
assessment.16,21

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                                     -13-
                                              Table 3
                                    COMPARATIVE  POTENCY  RANKINGS
                             MUTAGENESIS  TESTS
                              CARCINOGENESIS TESTS
                        AMES
SCE
L-5178Ya  BALE3
VIRAL
 ENHANCE   BAL8
TUMOR
 INITIATION
DIESEL: CATERPILLAR
NISSAN
OLDS
VW RABBIT
GASOLINE:
MUSTANG
COMPARATIVE
SOURCES :
CIGARETTE
COKE
ROO? TAR
STANDARDS :
B(a)P
4.3
100
23
22

25


7
18
7

1112
0
100
0
50

1


0
44
291

1750
1C
100
64
50

38


21
339
850

189
0
100
750
NTd

750


300
15
750

25000
0
100
25
50

50


200
800
2016

52000
0
100
0
NT

200


200
500
500

16700
0
100
28
6

16


0
355
120

16500
aln the presence of an Aroclor-1254  induced  rat  hepatic  S-9.
^Mouse skin tumor initiation in male and  female  sencar mice after  24  weeks  of  treatment.
cTesting incomplete at this time.
dNot tested.

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                                     -14-

Th e skin tumor initiation work is discussed later in this paper.

It  was  determined  that  the  Balb  test  is  not  suitable  for  a  complex
mixture^ so  that  the  Balb data cannot  be  used for any  comparisons.   Also,
there  is  some  concern  that  the  Ames  test  may  overreact  to  nitrogen
containing compounds  such as are  present  in Diesel particulates.   There  is
also concern  about whether bioassay test  results  in  general can  be  extra-
polated to human health effects.22

Nevertheless,  the  data  in Table  3 (especially the  skin tumor  initiation
data) can be used as an input for a Diesel risk assessment.

HERL has also done some work to  evaluate whether mutagens adsorbed on Diesel
particulates  are  released  in  the  presence  of  physiological  fluids.   This
work  showed   that  substantial  mutagenic activity  is  released from  Diesel
particles  upon incubation  with  serum  and  lung  cytosol.23   This work  is
continuing and  is  important in  determining  if physiological  fluids present
in the body alter the mutagenic activity of the particulate.

2.  Skin Tumorigenesis and Intratracheal Instillation

HERL has started long  term  projects  for skin tumorigenesis and intratracheal
instillation.

The skin tumorigenesis work was  done with motor  vehicle and other samples  as
discussed in  Section IVA1.  Both  C-57A and Sencar mice were  used.   Recent
results  with  the  C-57 black mice (a  relatively  non-sensitive breed)  were
negative (i.e.  no  carcinogenic  response) with  the  exception of one  dose  of
the roofing  tar samples.24  Work  with  the  more sensitive  Sencar  mouse does
show  a positive response  after  24 weeks  of treatment.25,26   These results
are given in  Table 3 above.  These results  are  preliminary ones; the results
from the full scale  study should  be available  within a  year.   OMSAPC feels
that  additional work  is  needed  for heavy  duty  Diesel samples.   Also,  the
cigarette smoke condensate showed no  result probably  due  to the  fact that
the active  components  are  too dilute   in the  condensate sample.   This work
should be repeated with a more active fraction of the condensate.

It  is  generally   thought^  that  the   skin  tumorigenesis  results  are  more
significant  from   a  human  health  viewpoint  than  bioassay  tests  since  the
whole animal  is involved.

The  intratracheal  instillation  work with  Syrian  golden  hamsters  involves
placing  specific  quantities  of  the   samples   listed   in  Table  3  on  the
trachea.   The doses  are repeated  at   certain  intervals.   The  advantage  of
intratracheal  instillation versus  inhalation  testing  is that  larger doses
can  be  administered  to  the  animals   in  a  very  controlled  fashion.   The
animals  are   then  followed over  their   lifetime  (about 2  years)  to  see  if
cancerous tumors develop.

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                                     -15-

This  work is  being  done under  an EPA Cooperative  Agreement  with  IITRI  in
Chicago.   Unfortunately, there  is  probably  no  way to  get  any  meaningful
results  until  the animals die since this  is  a lifetime study.  A  report  on
the work  is  scheduled for early  1983.

3.  Whole Animal  Inhalation

HERL-Cincinnati  has  done extensive  work  on  whole animal   inhalation  for
Diesel exhaust.   This work was described  in detail in the recent proceedings
of the EPA Diesel symposium held in 1979.

A  study2°>29  wag recently  completed  which  examined  the  effect   of  Diesel
particulate  exposure  on  the  ability of A/Jax mice to dispose  of radioactive
labelled  BaP that were  deposited  by  intratracheal   instillation.    While  it
was seen that mice exposed to Diesel exhaust  had  less capacity to  dispose of
the BaP, the significance of this  finding is not clear to OMSAPC.

Another animal inhalation program  involved  the  liver foci;  test  were run on
rats  that had most of their  livers surgically removed and  were fed  a special
choline-devoid or supplemented  diet for  3-6  months.  The  rats  were exposed
to either  clean  air or Diesel exhaust.   This test showed no  changes  in the
rats  exposed to Diesel exhaust.30

An initial study  was done with Strain A mice  which did  seem to show a slight
effect due  to  Diesel particulates  but  there were some questions  about  the
significance  of  the  results.^7,31   This  study  was   repeated  with  more
animals and the results are just being compiled now.

Another   study32   was   performed   with   hamsters.   A  sample  of  Diesel
particulate  extract  was intratracheally  instilled   in the   animals.   Lung
tissues  from  these   animals  were  later  analyzed  for  chromatid  exchange.
Recent analysis-^ shows  the  response from  this  test is linearly related  to
Diesel particulate  concentration.   Thus,  this  test   could  be  regarded  as  a
useful in-vivo test.   Future work  could involve using  other  samples includ-
ing more  Diesels  and possibly coke oven,  cigarette smoke, and roofing  tar
samples.

Another  project  that  is  currently  underway is intraperitoneal  injection  of
Diesel particulate,  coke oven,  cigarette  smoke condensate  and roofing  tar
samples  into the  lung cavities  of  laboratory  animals.   This  work will  be  a
supplement to the other work on these samples discussed  in Table 3.

Another  project-^ involved determining  if  Strain A mice  exposed  to  Diesel
exhaust had abnormal  sperm.  No  sperm abnormalities  were noted in  the Strain
A  mice.    Also,   the  urine  of   Swiss  mice  exposed   to Diesel  exhaust  was
analyzed  to  see   if it were  positive on  the  Ames test  due to  excretion  of
mutagenic  substances.35   No difference  was  found   in the   urine  of  mice
exposed to Diesel exhaust compared to the control group.

-------
                                     -16-

Finally,  HERL has  run  a  study^6  with  male  Chinese  hamsters  exposed  to
Diesel exhaust for  six months.  An  increase  in sperm abnormality was  noted
although no  increase  in chromosomal abnormalities in bone marrow  cells were
found  in  animals exposed  to  Diesel  exhaust.   Also, no  increase  in  sister
chromatid exchange was noted in bone marrow cells of these animals.

Other   Diesel   whole   animal   inhalation  work   is   underway  with   cats,
rabbits^?,  rats^° and  hamsters.    The  project  with cats  is  directed  more
towards non-carcinogenic lung effects.

OMSAPC  has  requested an  evaluation from  ORD on  the  general  usefulness  of
inhalation  studies  to  detect  increased  incidence  of  Diesel  cancer  in
laboratory  animals.   EPA wants  to  be certain  that  a negative  result   on  a
whole animal inhalation test is in fact significant.

B.  Carbon Monoxide Health Work

EPA  has  compiled a  detailed  summary  of health  effects  work  on  carbon
monoxide  in  the  recent Criteria  Document.3'   of particular interest to EPA
are  the numerous  studies^0*^ >42,43,44,45,46,47 which  show a  relationship
between carbon monoxide and the onset of  heart  pain in persons with  angina
pectoris.   This  work  has  been criticized  since  the  onset  of heart  pain can
be regarded  as  a subjective measure  that may not be  indicative  of further
heart  damage.   Also,  OMSAPC  has expressed   interest  in some  studies^°>^'
that have  related CO levels in the ambient  air  to  an increase in  hospital
admissions  due   to  cardiorespiratory complaints.   These studies  have  been
criticized  for  several reasons (basically a lack  of  showing  that the  CO
exposure  itself  was  responsible  for  the  health problems requiring  hospital
admission)  and  OMSAPC  has  expressed an  interest in  having them repeated.
These  and other  health  studies  have been   summarized  in   a  recent  review
article.50

Both OMSAPC  and   the  Office of Air  Quality  Planning  and Standards  (OAQPS)
have expressed a need  for  additional  CO  health  work.   ORD is  implementing
FY81 programs to meet these needs.

The  first  program is a replication of  the CO  clinical  studies relating  CO
exposure  to  actual changes  in heart parameters  during  exercise  for  people
with angina  pectoris.  This  work  will  be done with a radionuclide  gamma
camera  designed   to  determine  changes  in heart  parameters  (e.g.   chamber
discharge  volume  and  rates).    While   this  work   is  to  be   completed  by
December,  1982,  OMSAPC has requested a written progress report  in 1981.-^
Other CO work being done  by HERL  includes some clinical work to examine the
Coburn  equation   which  predicts  the  blood  carboxyhemoglobin  level  as  a
function of CO exposure.  This work will  be completed in  September  1982.

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                                     -17-

OMSAPC and OAQPS feel that work on  the  effect  of CO on susceptible subgroups
such as  fetuses  is needed.  HERL has started  some  work on pregnant  rats  to
determine if exposure to  CO  affects the newly born  rat  neonate.   An interim
report  on this  work which  is  now  underway  is  to be  prepared  later  this
year.   The  study will  be completed  in 1982.   HERL  also  plans  some  animal
studies  to determine the  effect of CO  alone  and in combination with  03  and
M>2  on   xenobiotic   metabolism  in  animals.   Additional  work   is   being
considered to  correlate  CO exposure and fetal carboxyhemoglobin  levels  with
cardiovascular and central nervous  system maldevelopment in the animal fetus
and neonate.   These  two projects would be complete in 1983.

HERL  has proposed  some  epidemiology feasibility  studies on  CO but  these
studies have not been  currently approved by the Mobile  Source  Research  Com-
mittee.  The one study would  determine  cardiovascular  effects  of CO exposure
at  high  altitude  while  the second  study  would  determine  the  effect  of
ambient carbon monoxide exposure on pregnancy outcome.

C.  Tier Bioassay Development

OMSAPC has need  for  a tier bioassay  test  for  both  the Section 211  fuel  and
fuel additive regulations  and the  Section  202  (a)(4) regulations  designed  to
assure that unregulated emissions  from motor  vehicles  do  not  cause  or  con-
tribute to an unreasonable risk to public health, welfare or  safety.   These
two sets  of  regulations  are being  currently  developed by OMSAPC.   The  tier
bioassay  test  would be  designed  to minimize the  cost  and   complexity  of
determining the  potential carcinogenicity of  these  emissions.   The  test  is
designed  in  a  tier  fashion  so  that  if  a  mixture  passes  the   first  test  (a
test  that should  result   in  few  false negatives),  no further   testing  is
needed.   If  it  fails, successive  tests of increasing complexity  are  needed
until  the mixture   passes  a test  (again  a  test   designed  for  few  false
negatives).    If a  mixture  fails  the  tier   bioassay process,  either  the
fuel/fuel additive for Section 211  or the vehicle for  Section  202 (a)(4) may
have to be altered until the emission products do pass the  tier tests.

A  problem with  developing  a  tier  test  is  to  specify   suitable  pass/fail
criteria for each successive test of  the tier.   HERL feels that they may not
be in a position to  specify these cut points.  Yet,  OMSAPC feels  that  HERL1 s
specifying  such cutpoints  is  an  inherent  part   in  developing  this  tier
approach.

The following tests  are being used by HERL:

                   1.   microbial mutagenesis
                   2.   cellular toxicity
                   3.   gene mutations in mammalian  cells
                   4.   mitotic  recombination in yeast
                   5.   sister chromatid exchanges in mammalian cells
                   6.   oncogenic transformation in  mammalian cells
                   7.   in  vivo/in  vitro  confirmatory  assays  employed  as
                        required.

-------
                                     -18-

Endpoints include:

              gene  mutations,  DNA damage,  chromosomal  effects,  oncogenic
              transformation   (carcinogenesis,   vn   vitro),   and   cellular
              toxicity.

A report  on  the short-term bioassay results and  a report on  correlation  of
short-term  bioassay test  results  is  due October 1981.  A revised  tier  will
be  constructed  and  investigated  with  a  final  tier  and  guidelines  being
available for OMSAPC in late  1982.  It is not known yet  how  toxic (i.e.  non-
cancer) emissions  will be handled  for Section  211 but  it  is possible  that
the approach that would be used for Section 202  (a)(4)  could  be  used here  as
well.  In  addition to  developing  the  tier biossay, HERL is  running limited
bioassay  tests  to  support ESRL for the  work  described in Section  II.   HERL
is also advising OMSAPC on the use of the Ames test for OMSAPC programs.
While  OMSAPC  now has  contractor  capability to  run  the Ames  test, HERL  is
still working on data  analysis  for the Ames test.  This  capability  is  in the
process of being transferred  to OMSAPC.

V.  Diesel Carcinogenic Risk Assessment

The  major output  of   the  massive  ORD Diesel  research  program  is  to  be  a
revised  carcinogenic   risk assessment  projecting the  potential  number  of
cancer cases that  could be associated  with  exposure  to light  and heavy  duty
Diesel exhaust.

An  initial  risk assessment^ was  based on  assuming  the  potency  of  Diesel
exhaust extract  is equivalent  to  that  of  the benzene-soluble organics  from
coke oven emissions.   The best estimate  number  of projected  cancer cases  in
1990  would  be  about  350   and   670  for  light  and   heavy  duty  Diesels
respectively.   Since  this   risk  assessment  was  released,  a number  of
criticisms of  it have been made  by  various  people  including  the National
Academy of Sciences.

It  is  agreed that  a  new  risk assessment  is  needed.   ORD  is  scheduled  to
release a new  risk  assessment shortly.   The  biggest  change  that   could  be
expected  in the new risk  assessment is that a lower potency  would be assumed
for Diesel exhaust compared to coke  oven emissions based on  the recent  skin
tumorigenesis results.25,26,54

ORD  did  release a report recently of great  value to  OMSAPC on the  London
Transit Worker  study.54   This  epidemiology study  of  London  transit workers
exposed to Diesel  bus emissions has  been  cited by many  people as  a  strong
indication  that Diesel   emissions  result  in  no  excess cancer risk.   ORD
analyzed  the  study  parameters statisically  looking  at  the  particle  level
exposure  time,  and number of  people involved.   It was  their  conclusion  that
thousands  of  excess  cancer   deaths   could  result   in  the  United  States
population as a whole  and  still be consistent with the  results of the London
Transit Worker  study.   The ORD  analysis has  been reviewed  both inside  and
outside EPA by various experts.

-------
                                     -19-

VI  Health Effects Institute

In  late  1980,  the  Health  Effects  Institute  was  formed  to conduct  health
research for mobile source  emissions.  While  the  Institute is jointly funded
by the industry and  EPA,  it is  set  up  as an  independent  entity  so  that  the
research done will be purely scientific and free of any bias.

At  this  point,  the Health  Effects  Institute  is  formulating plans   for  the
type  of   research they  would  implement.   EPA  and   the  industry  will  be
specifying  the   type  of  work  that  is  important  from their  perspectives.
Specific projects should be started later this year.

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                                     -20-

REFERENCES

1.  Penny Carey  and Janet Cohen,  EPA,  "Comparison of Gas  Phase Hydrocarbon
    Emissions  from  Light Duty  Gasoline  Vehicles  and  Light-Duty  Vehicles
    Equipped   with   Diesel   Engines",   EPA-OMSAPC   Report   (CTAB/PA/80-5),
    September 1980.

2.  Frank  Black  and  Lisa  Doberstein,  EPA,   "Filter Media  for  Collecting
    Diesel Particulate Matter",  ESRL Internal Report to be  submitted  to the
    Society of Automotive Engineers  for publication, March 1981.

3.  Federal   Register,   "Standard   for   Emission   of   Particulate   for
    Diesel-Fueled Light-Duty  Vehicles  and Light-Duty Trucks",  Vol. 45,  No.
    45, 14496-14525, March 5, 1980.

4.  Dennis Schuetzle, Frank S. C.  Lee,  and Thomas  J.  Prater,  Ford Motor Co.,
    and  Silvester  B.  Tejada,   EPA,   "The   Identification   of  Polynuclear
    Aromatic  Hydrocarbon  Derivatives  in  Mutagenic Fractions  of  Diesel
    Particulate  Extracts",  presented  at  the  10th Annual  Symposium  on  the
    Analytical Chemistry  of  Pollutants, Dortmund,  Germany,  May  28-30,  1980
    and  to  be  published in the  International  Journal  of  Environmental
    Analytical Chemistry.

5.  Peter A. Gabele, Frank M. Black, Foy G. King,  Roy B.  Zweidinger, and Rex
    A. Brittain,  EPA,  "Exhaust  Emission Patterns  from Two  Light-Duty  Diesel
    Automobiles",  Society of  Automotive  Engineers  Paper  810081,  February
    1981.

6.  Richard  E.  Gibbs,  James  D.  Hyde,  Stanley  M.  Byer,  New  York  State
    Department  of  Environmental  Conservation,  "Characterization  of  Par-
    ticulate Emissions  from  In-use  Diesel  Vehicles", Society  of Automotive
    Engineers Paper 801372, October  1980.

7.  Harry E. Dietzmann, Mary Ann Parness,  Southwest  Research  Institute,  and
    Ronald L. Bradow, EPA, "Emissions  from Trucks  by Chassis Version of 1983
    Transient  Procedure", Society  of  Automotive  Engineers  Paper  801371,
    October 1980.

8.  J. Lewtas  Huisingh,   David L. Coffin,  Ronald  L.  Bradow,  Larry Claxton,
    Ann  Austin,   Roy  Zweidinger,  Robert  Walter,  Joe Sturm,  and  Robert  H.
    Jungers,  "Comparative Mutagenicity  of Combustion  Emissions  of  a  High
    Quality No. 2 Diesel  Fuel Derived  from Shale Oil  and  a Petroleum Derived
    No. 2 Diesel  Fuel",  Interagency Symposium on  the Health  Effects Invest-
    igation of Oil Shale Development, Gatlinburg, Tennessee, June 1980.

9.  James L. Repace,  Wayne R. Ott,  and Lance A.  Wallace, EPA,  "Total Human
    Exposure  to  Air  Pollution",  Air   Pollution   Control  Association  Paper
    80-61.6, June 1980.

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                                     -21-

10. Mark Wolcott,  EPA,  "Summary  Report  of  Several  Ambient Carbon  Monoxide
    Studies", EPA-OMSAPC Report (TEB-81-9), November 1980.

11. William F. Biller,  Thomas  B.  Feagans,  Ted R. Johnson,  George  M.  Duggan,
    and James E.  Capel, "Estimated Exposure to Ambient  Carbon Monoxide Con-
    centrations   Under   Alternative   Air   Quality   Standards",   EPA-OAQPS
    Strategies and  Air Standards  Division  Draft  Report,  RTP, N.C.  27711,
    August 1980.

12. Lance  Wallace,   EPA,   "Personal  Air   Quality  Monitors:  Past  Uses  and
    Present Prospects", ACS  Proceedings,  4th Joint Conference  on  Sensing of
    Environmental Pollutants, 1978.

13. A. C.  Gullon,  Environment Canada  "Motor Vehicle  Pollutants in  Canadian
    Cities", Air Pollution Control Association Paper 80-2.1, June 1980.

14. Anthony D.  Cortese  and  John  D.  Spengler,  "Ability of  Fixed  Monitoring
    Stations to Represent  Personal Carbon  Monoxide Exposure",  Journal  of the
    Air Pollution Control  Association 26,  1144, 1976.

15. James  W.  Jabara  and   Thomas  J.  Keefe,  "Carbon  Monoxide:  Dosimetry  in
    Occupational  Exposures in Denver  Colorado",  Archives  of  Environmental
    Health, _35,  198, 1980.

16. Joseph H.  Somers,   EPA,  "Trip  Report   on EPA-ORD  Diesel Risk  Assessment
    Meeting", OMSAPC memo,  March 4, 1981.

17. Larry  Claxton  and  Joellen  L.  Huisingh,  EPA,  "Comparative  Mutagenic
    Activity  of  Organics  from  Combustion  Sources",   Proceedings  of  the
    Symposium on Pulmonary Toxicology of Respirable Particles,  March 1980.

18. J. L.  Huisingh,  R. L.  Bradow,  R.   H.  Jungers,  B.  D.  Harris,  R.  B.
    Zweidinger,  K. M. Gushing, B.  E. Gill, and R. E. Albert,  "Mutagenic and
    Carcinogenic   Potency   of  Extracts  of  Diesel and  Related  Environmental
    Emissions:    Study   Design,    Sample   Generation,    Collection,    and
    Preparation", EPA-HERL Report, February 1981.

19. J.  Lewtas Huisingh,   EPA,  "Short-term  Carcinogenesis  and  Mutagenesis
    Bioassays of Unregulated Automotive Emissions", Bulletin of the  New York
    Academy of Medicine, in press 1981.

20. Bruce  C.  Casto,  George  G.  Hatch,  Shiu  L. Huang,  Joellen L.  Huisingh,
    Stephen Nesnow,  Michael  D. Waters, "Mutagenic  and  Carcinogenic  Potency
    of Extracts  of  Diesel  and  Related  Environmental   Emissions;  In  Vitro
    Mutagenesis  and Oncogenic  Transformation",  EPA-HERL  Report,  February
    1981.

21. Jeff Alson,  EPA, "Trip Report  on the Diesel  Health  Effects Workshop Held
    in RTP on February 24-25, 1981",  OMSAPC memo, March  1981.

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                                     -22-

22. "Health Effects of Exposure  to  Diesel  Exhaust",  the report of the Health
    Effects Panel  of  the Diesel  Impacts Study Committee,  National  Research
    Council, National Academy of Sciences, 1980.

23. Leon  C.  King,  Mike  J.  Kohan,   Ann  C.  Austin, Larry D. Claxton,  and J.
    Lewtas Huisingh, EPA, "Evaluation of the Release of Mutagens from Diesel
    Particles  in the  Presence of Physiological Fluids",  Environmental Muta-
    genesis, in press 1981.

24. Stephen Nesnow,  EPA, "Report  on  Skin  Tumorigenesis  Studies of  Diesel
    Emissions and Related Samples on  C57 Black Mice", ORD-HERL memo, January
    12, 1981.

25. Stephen Nesnow, EPA, "Report on Skin Tumorigenesis Studies of Volkswagen
    Turbo  Rabbit  and Home  Heater  Samples on  Sencar Mice",  ORD-HERL memo,
    January 24, 1981.

26. Stephen Nesnow, Larry  L. Triplett,  and  Thomas J.  Slaga,  "Tumorigenesis
    of  Diesel  Exhaust,   Gasoline  Exhaust,  and  Related Emission  Extracts on
    Sencar Mouse Skin",  ORD-HERL Report, 1981.

27. "Health Effects of  Diesel Engine Emissions:  Proceedings of  an Internat-
    ional  Symposium", EPA  Report 600/9-80-057ab,  Volumes  1 and  2,  November
    1980.

28. E. T.  Cantrell, H. W. Tyner, W. B.  Peirano,  and  R.  M.  Danner,  "Benzo(a)
    pyrene  Metabolism  in Mice  Exposed to  Diesel  Exhaust:   Metabolism  and
    Excretion", Environment  International, in press 1981.

29. H. W. Tyner, E. T.  Cantrell, R.  Homes,  I.  P.  Lee,  W.  B.  Peirano,  and R.
    M.  Danner,  "Benzo(a)   pyrene  Metabolism  in  Mice  Exposed  to  Diesel
    Exhaust: Uptake  and Distribution",  Environment  International,  in press
    1981.

30. M.A.  Pereira,  H.  Shinozuka,  and B.  Lombardi,  "Test  of  Diesel  Exhaust
    Emissions  in the Rat Liver Foci  Test", ORD-HERL Report, 1980.

31. John G. Orthoefer, Wellington Moore, Dale Kraemer,  Freda  Truman,  Walden
    Crocker, You Yen Yang, EPA,  "Carcinogenicity  of  Diesel  Exhaust  as Tested
    in Strain A Mice",  ORD-HERL Report,  1980.

32. Donald  E.   Rounds,   "Mutagenicity   of  Diesel  Exhaust  to  Hamster  Lung
    Tissue", ORD-HERL Report, 1980.

33. William E.  Pepelko, EPA, Letter  to  Gerald  Rausa  on Sister  Chromatid
    Exchange,  March 10,  1981.

34. M. A.  Pereira, P.  S.   Sabharwal,   L.  Gordon, and A.  J.  Wyrobek,  "The
    Effect of Diesel Exhaust  on  Sperm-shape  Abnormalities  in Mice",  ORD-HERL
    Report, 1980.

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                                     -23-

35. M.  A.  Pereira, T.  H.  Connor,  J.  Meyne, and  M.  S.  Legator,  "Metaphase
    Analysis,  Micronuclei  Assay  and   Urinary  Mutagenicity  Assay  of  Mice
    Exposed to Diesel Emissions", ORD-HERL Report, 1980.

36. M.  A.  Pereira, P.  S.  Sabharwal,  P. Kaur,  C.  B.  Ross,  A.   Choi,  and T.
    Dixon,  "In-vivo  Detection  of  Mutagenic Effects  of  Diesel  Exhaust  by
    Short Term Mammalian Bioassay", ORD-HERL Report, 1980.

37. "Study  of  the  Teratologic  Effects  of Long-term   Exposure  to  Diesel
    Exhaust   (Rabbits)",   Final   Report  by   Wil  Research   Laboratories,
    Cincinnati, Ohio  for EPA Contract 68-03-2652, 1980.

38. "Study  of  the  Teratologic  Effects  of Long-Term   Exposure  to  Diesel
    Exhaust   Emissions   (Rats)",   Draft  Final   Report   by   Wil  Research
    Laboratories,  Cincinnati, Ohio  for EPA Contract 68-03-2652, 1980.

39. "Air  Quality  Criteria  for  Carbon   Monoxide  (Preprint)",   EPA  Report  -
    600/8-79-022,  CO  Criteria  Document, Office of  Research  and Development,
    October 1979.

40. E. W. Anderson, R.  J.  Andelman, J.  M.  Strauch, N.  J.  Fortuim,  and J. H.
    Knelson,  "Effects of  Low-Level Carbon Monoxide on Onset and Duration of
    Angina  Pectoris:  A Study  on 10 Patients  with Ischemic Heart Disease",
    Ann. Intern. Med. _7£, 46-50, 1973.

41. W. S. Arnow, "Effect of  Passive Smoking  on  Angina  Pectoris", N.  Engl. J.
    Med. 299. 21-24,  1978.

42. W.  S.  Aronow  and  J.  Cassidy,  "Effect  of  Carbon  Monoxide  on Maximal
    Treadmill Exercise:   A Study  in Normal  Persons",  Ann. Intern.  Med.  83,
    496-499,  1975.

43. W. S. Aronow,  J.  Ferlinz,  and F. Glauser, "Effect of Carbon Monoxide on
    Exercise  Performance  in  Chronic Obstructive Pulmonary Disease",   Am. J.
    Med. j>3,  904-908, 1977.

44. W.  S.   Aronow  and  M.W.   Isbell,  "Carbon  Monoxide  Effect   on Exercise-
    Induced Angina Pectoris", Ann.  Intern. Med.  79, 392-395,  1973.

45. W. S. Aronow,  C.  N.  Harris, M.  W.  Isbell,  S. N. Rokaw, and B. Imparato,
    "Effect  of  Freeway  Travel  on  Angina  Pectoris",  Ann.  Intern.  Med.  77,
    669-676,  1972.

46. W.  S.  Aronow,  J. Cassidy,  J.  S. Vangrow,  H.  March,  J.  C. Kern,  J.  R.
    Goldsmith,  M.  Khemka, J.   Pagano,   and M.  Vawter,  "Effect  of Cigarette
    Smoking and  Breathing  Carbon Monoxide on Cardiovascular Hemodynamics on
    Anginal Patients", Circulation  50,  340-347, 1974.

47. W. S. Aronow,  "Effect  of 2% Venous  Carboxyhemoglobin on  Exercise-Induced
    Angina Pectoris", 1980.

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                                     -24-

48. T. L. Kurt,  R.  P.  Mogielnicki, and  J.  E.  Chandler, "Association  of the
    Frequency of  Acute  Cardiorespiratory Complaints  with  Ambient  Levels  of
    Carbon Monoxide", Chest J4, 10-14, 1978.

49. T. L. Kurt,  R.  P. Mogielnicki,  J.  E.  Chandler,  and K.  Hirst,  "Ambient
    Carbon  Monoxide  Levels   and  Acute  Cardiorespiratory   Complaints:   An
    Exploratory Study",  Am. J. Health 69_, 360-363, 1979.

50. Gerard M.  Turino,  "Effect  of  Carbon Monoxide  on  the  Cardiorespiratory
    System",   Journal  of  the  Air  Pollution Control  Association,   63,  253A,
    1981.

51. Mathew L. Petrovick, EPA,  "Biomedical Instrumentation and Techniques for
    Cardiovascular  Carbon  Monoxide  Exposure   Studies",   ORD-HERL  Report,
    August 27, 1980.

52. Roy E. Albert, EPA, "Initial Review  of  the  Potential Carcinogenic  Impact
    of Diesel Exhaust",  EPA Memo, June 11, 1979.

53. Todd W.  Thorslund, EPA, "A Suggested  Approach  for the Calculation  of the
    Respiratory Cancer Risk Due to Diesel  Engine Exhaust",  EPA-ORD Report,
    February 1981.

54. Todd  Thorslund,  EPA,  "Answer  to  the  Posed Question  'Are the  results
    obtained  in  the London Transit  Worker  Study  sufficient  to dismiss  any
    concern regarding the  potential  cancer hazard  for the  U.  S,  population
    in the  future,  due  to Diesel engine exhaust?"1, EPA-ORD  memo,  January
    29, 1981.

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