ASSOCIATION OF SMOKING WITH CHRONIC DISEASES
1. CANCER
MECHANISMS BEGIRNING TO BE UNDERSTOOD
BIOASSAYS AVAILABLE
A) MUSE SKIN PAINTING
B) INHALATION TESTS
c) SHORT-TERM TESTS
2. CARDIOVASCULAR DISEASES
MECHANISMS POORLY UNDERSTOOD
NO SUITABLE ANIMAL MODEL
3, CHRONIC OBSTRUCTIVE LUNG DISEASES
MECHANISMS POORLY UNDERSTOOD
NO SUITABLE LABOPATORY MODEL
9MZ8601

CANCER - POSSIBLE MECHANISMS
1, EXISTENCE OF A RELATIOUSHIP BETWEEN
CARCIROGENESIS AND MUTAGERESIS
2. THE MUTATION THEORY OF CANCER -
CANCER RESULTS FROM MUTATION OF SOMATIC CELL
3. MUTATED SOMATIC CELLS OUTGROW
SURROUNDING CELLS
4. LOUG PERIOD BETWEEN MUTATIONAL EVE11T AND
DEVELOPMENT OF CANCER
5. TWO-STAGE THEORY
A) INITIATION
B) PROMOTION
MOZ8601

EVOLUTION OF SHORT-TERM MICROBIAL TESTS
CRICER - SOMATIC CELL MUTAT104
CARCINOGENS SHOULD BE MUTAGENS
METABOLISM OF SOME CHEMICALS TO ACTIVE FORM
CLOSELY RELATED FORMS SHOW DIFFERENT ACTIVITIES
CARCINOGEN TESTING - ANIIIALS
MUTAGEN TESTING - MICROORGAINISMS (FiEHEIRALLY)
INCORPORATION Of MAMMALIAN ACTIVATING EMZYMES
IN MICROBIAL TESTS
ANIMAL CARCINOGENS (MOST) ARE MICROBIAL MUTAGENS
IDENTIFICATION OF MUTAGENS PER SE IMPORTANT III
PROTECTING HUMAN GENE POOL
SAFETY-REGISTRATION COMMITTEES WORLD-WIDE ACCEPT THE
AMES TEST FOR MUTAGENIC, EVALUATION OF CHEMICALS AND
CONSIDER IT IMPORTANT IN CARCINOGENICITY TESTING,
91POI8601

tz
n
0
TOBACCO SMOKE ARCINOGENICITY H.C.I., U.SJ, STUDIES
2 FACTORS ASSOCIATED WITH
Z.
REDUCED ACTIVITY NO CHANGE INCREASED ACTIVITY
> 1. HIGH POROSITY PAPER 1. WIDTH OF TOBACCO CUT 1. FILTERS
0
w (CITRATE) A) CELLULOSE ACETATE
>I
n 2. TOBACCO STEM 9 EXPANDED 2. PAPER POROSITY B) PERMANGANATE
n
0
3. RECONSTITUTED SHEET 3. RECONSTITUTED SHEET 2. RECONSTITUTED SHEET
A) RTS PAPER PROCESS A) RTS SLURRY PROCESS A) SLURRY PROCESS
0
B) WATER EXTR PAPER PROCESS B) PAPER PROCESS WITH B) PAPER PROCESS - EXTRACTED
WITHOUT ADDITIVES WATER 65%; INORGANIC, 25%;
z 4. LOW NICOTINE TOBACCO ADDITIVES, 101
4. EXPANDED TOBACCO
C) PAPER PROCESS - NORMAL
5. EXPANDED TOBACCO 5. REYNOLDS PUFFED TOBACCO NICOTINE
A) PHILIP MORRIS PROCESS 6. SUGAR
3. ARTIFICIAL TOBACCO SUBSTITUTE
B) FREEZE DRIED 7. HUMECTANT ATS-B
6. ARTIFICIAL TOBACCO
8. ADDITIVES
SUBSTITUTES - ATS-A A) ZINC OXIDE
7. ADDITIVES B) COCOA
A) POTASSIUM NITRATE 9. SUCKERING AGENTS
B) MAGNESIUM NITRATE FATTY ALCOHOL
8. -FILTER VENTILATION
00L8601

A SUMMAR" IF B.A.T. MUTAGENICITY STUDIEF
FACTORS ASSOCIATED WITH
REDUCED ACTIVITY HO CHANGE IMCREASED ACTIVITY
1. LOWER CIRCUMFERENCE 1. PUFF DURATION 1. FILTER VENTILATION
2. PAPER POROSITY 2. PUFF FREQUENCY 2. HIGHER FILTER PRESSURE DROP
3. HIGHER PUFF VOLUME 3. WIDTH OF TOBACCO CUT 3. HIGHER TOBACCO MOISTURE
4. VIRGIIIIA TOBACCO 4. BUTT LENGTH 4. BURLEY TOBACCO
5. SHEET MATERIALS 5. FILTERS 5. DARK TOBACCO
A) GERLACH A) HEM111 6. TOBACCO PROTEIN
B) PCL 8) IONEX
c) SCHWEITZER 7. NITPATE
D) CYTREL 6. ANNULAR CIGARETTES
7. NICOTINE 8. NICOTINE
6. STEM 9. CASINGS
8. MERTHOL
7, EXPANDED TOBACCO 10, DITHIOCARBAMATE
8. REDUCING SUGAR 9. COCOA (MANEB) APPLICATION
9. CASINGS
10. BUTTERFAT
11. KRETEK
0DOZ8601

0
TOBACCO SMOKE CARL OGENICITY - B.A.T. - SUPPORTE STUDIES
z
z FACTORS ASSOCIATED WITH
REDUCED ACTIVITY NO CHANGE INCREASED ACTIVITY
> 1. LOWER CIRCUMFERENCE 1. REDUCING TOBACCO MOISTURE 1. PAPER POROSITY
__4
0
2. HIGHER PUFF VOLUME. 2. GRANULATED TOBACCO 2. SILICA GEL FILTER
n
3, INCREASED FLOW RATE 3. FILTERS 3. INCREASING TOBACCO MOISTURE FROM
A) ACETATE 12% To 20%.
4. INCREASED CUTS PER B) POLYETHYLENE GLYCOL
0 INCH 4. DARK TOBACCO - GERMAN
> 0 CHARCOAL
I 5. SUN-CURED - INDIAN
5 5. BURLEY TOBACCO AND D) CHARCOAL (PELLET TYPE)
z STEM E) CELLULOSE ACETATE 6. CIGAR LEAF
6. VIRGINIA STEM. 4. ADDITIVES 7. GRANULATED IN CIGAR
7. RECONSTITUTED SHEET A) MALIC ACID 8. RECONSTITUTED SHEET
A) GERLACH B) 2% DIETHYLENE GLYCOL A) ARENCO
B) PCL 0 1Z GALLIC ACID-N- B) BORGWALDT
0 SRT PROPYL ESTER 0 AMF
D) SWITZER 9. ADDITIVES
8. ADDITIVES A) POTASSIUM MALATE
A) POTASSIUM NITRATE B) POTASSIUM HYDROGEN MALATE
B) COPPER NITRATE 0 COPPER POTASSIUM MALATE
c) POTASSIUM NITRITE D) POTASSIUM CARBONATE (NO STEAM)
D) SODIUM NITRATE E) NIMONIUM SULPHAMAtE
E) POTASS I UM CARBONATE
(PLUS STEAM)
MUM[

A COMPARISON OF BIOLOGICAL ACTIVITIES OF CONDENSATE
IN THE CARCINOGENICITY AND MUTAGENICITY TESTS
CONCLUSIONS DRAWN FROM TESTS FOR WHICH THERE WAS
AGREEMENT DISAGREEMENT
LOWER CIRCUMFERENCE PAPER POROSITY
FILTER VENTILATION VIRGINIA TOBACCO
HIGHER PUFF VOLUME BURLEY TOBACCO
EXPANDED STEM
WIDTH OF TOBACCO CUT (0) CASINGS
HIGHER TOBACCO MOISTURE (4-)
DARK TOBACCO (+)
EXPANDED TOBACCO
GERLACH SHEET
PCL SHEET
CYTREL (-)
SCHWEITZER SHEET f%-)
VIRGINIA STEY, (-)
NICOTINEE ADDITION
COCOA ADDITION (0)
cx:)
CC)
BAT Co LTD MINNESOTA TOBACCO LITIGATION

A COMPARISON OF THE JANUS SERIES IN
CARCINOGENICITY AND MUTAGENICITY TESTS
EFFECT ON
CIGARETTES CARCINOGENICITY MUTAGIENICITY
B9 SERIES
ETHANOL EXTRACTION - -
E- 'Al"l'OL EXTRACT RETURNED 0 0
PCL PLUS GERLACH ADDITIVE - -
GERLACH SHEET - -
GERLACH SHEET FROM ETHPAOL - -
EXTRACTED TOBACCO
B11 SERIES
PCL - -
S RT
B.LD SERIES
INCREASED PAPER POROSITY
INCREASED PRESSURE DROP O/+
FILTER TIP VENTILATION 0 0
ELECTROSTATIC PERFORATION
PAPER ADDITIVE 0 0
L
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BAT Co LTD MINNESOTA TOBACCO LITIGATION

A COMPARISON OF JANUS B15 SERIES CIGARETTES IN THREE BIOLOGICAL TESTS
BIOLOGICAL ACTIVITY
CIGARETTE 8 WEEK
PARAMETER INHALATION MOUSE SKIN AMES
STUDIED STUDY CARCINOGENICITY MUTAGENICITY
PAPER INCREASE INCREASE INCREASE ASSOCIATED
Pr )s I TY ASSOCIATED WITH ASSOCIATED WITH WITH DECREASE IN
DECREASE IN INCREASE IN ACTIVITY.
ACTIVITY. ACTIVITY,
FILTER INCREASE (A) PAPER TYPE INCREASE ASSOCIATED
PRESSURE ASSOCIATED WITH T6_5Z- WITH INCREASE IN
DROP DECREASE IN NO EFFECT ACTIVITY.
ACTIVITY.
(B) PAPER TYPE
9 RI 9-5 C
INCREASE ASSOCIATED
WITH INCREASE IN
ACTIVITY.
FILTER TIP NO EFFECT NO EFFECT NO EFFECT
VENTILATION
ELE CT RO S TA T I C ELECTROSTATIC ELECTROISTATI C
PERFORATION PERFORATION PERFORATION
ASSOCIATED WITH ASSOCIATED WITH
INCREASE IN INCREASE IN
ACTIVITY ACTIVITY
PAPER ADDITIVE NO EFFECT NO EFFECT
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-C:b.
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BAT Co LTD MINNE@OTA TOBACCO LITIGATION

A COMPARISON OF THE JANUS B11 SERIES CIGARETTES IN SEVERAL BIOLOGICAL AND BIOCHEMICAL TESTS
z
z
>
-i PROMOTION MOUSE SKIN SEBACEOUS GLAND - AMES
0
w CIGARETTE Z AN114ALS BEARING TUMORIGENICITY BOSE TO PRODUCE 50ff MUTAGENICITY flMFI
>
n IIYPERPLASIAS AND ANIMALS SUPPRESSION TEST
n
0 NEOPLASIAS SHOWING TUMOURS
B11-1 24 (3 )A 4G (3)A LI 2. 2 (3 )A 3.83 (3 )A G IO (3)A
0 (CONTROL)
z
B11-2 23 MA 39 (41.) 1 211.9 (2 )A 3.44 (2)" 4.5 (2)B
(PCL)
B11-3 17 UP 17 (1)c 39.9 M' 3.42 (1)" 4.3 (1)"
(SRT)
THE NUMBERS GIVEN' III PRACKIETS ARE CIGARETTE RANKINGS.
C011DENSATE ACTIVITIES FOLLOWED BY A DIFFERENT LETTER
ARE S I Gil I F I CANTU D I FFERENT (p ( 0. 05)
SDOL8601

tz
n A COITARISON OF THE JANUS bi SERIES 13 SEVERAL BIOLOGICAL iii'D BIOCHEMICAL TESTS
0
z CIGARETTE CARCINOGENICITY
z CODE AMES Z TUMOURS
AND DESCRIPTION MUTAGEHICITY DOSE MG
>
4 40 60 x
0
w
n D9-1: FLUE-CORED BLEND 1.03 + 0.19 (6)A 66 (6) 78 (6) 72 (6)A
(LAMINA/CRS, 60/40)
0 B9-2: ETHANOL EXTRACTED 0.90 i 0.18 (4)' 50 (4) 57 (4) 51 (4 )B
)A
0 B9-3: ETHANOL EXTRACT 1.12 + 0.23 (6 60 (5) 60 (5) 60 (5)c
z RETURNED
B9-4: MONTREAL PCL 0.57 1 0.13 (3)' 42 (3) 45 (3) 48 (3)B
PLUS GERLACH ADDITIVE
B9-5: GERLACH SHEET 0.36 0.10 (I)D 7 (2) 38 (2) 31 (2)D
B9-6: GERLACH SHEET 0.48 0.13 (2)c 19 (1) 25 (1) 18 (1)E
FROM ETHXNOL EXTRACTED
TOBACCO
THE NUMBERS GIVEN IN BRACKETS ARE CIGARETTE RANKINGS.
BIOLOGICAL ACTIVITIES FOLLO!!ED BY A DIFFE.RENT LETTER ARE
9DOZ9601 111111FICAHTLY DIFFERENT (P4 0,95)1



CIGARETTES TESTED IN SKIN CARCINOGENICITY AND
BACTERIAL MUTAGENICITY TESTS - SOME DIFFERENCES
JANUS CONCLUSION FROM
SERIF$ . . . AMES MU TAG EN IC I TY SKIN CARCINOGENICITY
GERLACH SHEET FROM FLUE-CURED GERLACH SHEET FROM ETHANOL
Tin TOBACCO SHOWED THE -LOWEST EXTRACTED TOBACCO SHOWED
ACTIVITY. THE LOWEST ACTIVITY.
B, I I NO DIFFERENCE SEEN SIGNIFICANT DIFFERENCE SEEN
BETWEE14 PCL AND SRT BETWEEN PCL AND SRT.
INCREASE IN PAPER POROSITY INCREASE IN PAPER POROSITY
B15 ASSOCIATED WITH DECREASE ASSOCIATED WITH INCREASE
IN ACTIVITY IN ACTIVITY,
INCREASE IN FILTER INCREASE IN FILTER
PRESSURE DROP ASSOCIATED PRESSURE DROP ASSOCIATED
WITH INCREASE IN WITH:
ACTIVITY. 0) NO CHANGE (PAPER TYPE
3650
00INCREASE IN ACTIVITY
(PAPER TYPE DM 950
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BAT Co LTD I TOBACCO LITIGATION

TOBACCO SMOKE CARCINOGEhICITY IN INHALATION TESTS
LARYNGEAL TUMOURS IN HAMSTER
1. DONTENWILL ET AL., Z. KREBSFORSCH, 89, 153, 1977
100% TOBACCO CIGARETTES HIGHER IN ACTIVITY THAN
50-50Z BLEND OF CYTREL AND TOBACCO.
2. BERNFELD ET AL., J. NATL. CANCER INST., 63, 675, 1979.
1002 TOBACCO PRODUCED LARY14GEAL TUMOURS,
100% CYTREL DID NOT PRODUCE TUMOURS
CXD
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BAT Co LTD MINNE@OTA TOBACCO LITIGATION

tz
SUMMARY OF BIOLOGICAL RESEARCH AT McGILL UNIVERSITY
1. SMOKE INHALATION STUDIES USING LABORATORY ANIMALS AND THE
z B.A.T.-WON SMOKING MACHINE.
2. BIOCHEMICAL END POINTS:
> A) INDUCTION OF ARYL HYDROCARBON HYDROXYLASE
i
0
tz (POSSIBLE ROLE IN CANCER).
n
B) ABILITY OF TISSUE TO ACTIVATE PROCARCINOGENS
IN THE AMES SALMONELLA SYSTEM,
0
3. AHH NOT INDUCED IN GUINEA PIG LUNG
KIDNEY RESPONDS
z
4. GUINEA PIG KIDNEY UNIOUE - INDUCTIOn OF AHH PARALLELS
METABOLIC CAPABILITY4
5. RAT LUNG HIGH SENSITIVE - AIIH INDUCED BY DILUTED SMOKE
FROM A 1 MG CIGARETTE,
6. AMBIENT SMOKE INDUCES AH111 IN PAT LUNG$
7. EFFECTS OF SMOKE EXPOSURE ON PERINATAL ANIMALS,
MILK AND PLACENTAL TRANSFER OF SMOKE COMPONENTS
IN RAL BUT NOT IN GUINEA PIG AND HAMSTER.
14 DAY RAT PUP LIVER VERY SENSITIVE TO SMOKE EXPOSURES
690L8601

FUTURE STUDIES
1. EXPLORE THE SHORT TERM TESTS, INCLUDING TESTS FOR
ffPROMOTION" ACTIVITY OF TOBACCO SMOKE.
2. STUDY PROTECTIVE MECHANISMS IN ANIMAL MODELS.
3. USE OF AMES TEST TO STUDY:
A) TOBACCO TYPES, PARTICULARLY DIFFERENT BURLEYS.
B) SMOKE FRACTIONATION.
c) ANTIMUTAGENS.
D) ROLE OF NICOTINE.
E) DESIGN AND TEST 'LOW ACTIVITYO CIGARETTES.
4. USE OF AMES TEST TO MONITOR:
A) RECONSTITUTED TOBACCOS.
B) ADDITIVES - FLAVOURINGS, CASINGS, ETC.
c) NEW MARKET ENTRIES.
co
BAT Co LTD I T013ACCO LITIGATION

03
z MUTAGENICITY AND CARCINOGENICITY (INTERIM) OF JANUS B14 CIGARETTES
z
0
CIGARETTE CODE AND MUTAGENICITY - SPECIFIC CARCINOGENICITY TUMORIGENIC
n TNA IN CONDENSATE ACTIVITY-NO. OF HIS+ RATIOS (WEEK 100)+
(MG) REVERTANTS OF TA-98 PER
PG CONDENSATE
> A 1. 00 (1) A
B14-1 - - - - 0.56 2.05 i 0.31 (1)
;5
z B14-2 - - - - 0.80 2.07 + 0.110 (2) 1.09 (2) A
B14-3 - - 1.22 2.29 0.46 (3) 1. 23 (3) A
B14-4 - - - - 1,62 2,53 0.53 (4) c 1.69 (4) B
PLCK-20 - - - 1.70 2,11 0,40
THE NUMBERS GIVEN III BRACKETS ARE RANKS.
BIOLOGICAL ACTIVITIES FOLLMED BY A DIFFERENT
LETTER ARE SIGNIFICAMLY DIFFrRENT (p c n-05).
16VOL8601

FUTURE-STUDIES
EXTERNAL - (INDUSTRY SPONSORED?)
1. SMOKING AND I [II UN MECHANISMS.
2. PROTECTIVE MECHANISMS - ANTICANCER ACTIVITY
OF CHEMICALS IN ANIMAL MODELS.
3, IDENTIFICATION OF SUSCEPTIBLE INDIVIDUALS.
4. DUBOW'S BIOTECHNOLOGY APPROACH TO TOXICOLOGY.
co
BAT Co LTD MINNESOTA TOBACCO LITIGATION

z DUBOW STUDIES WITH CIGARETTE SMOKE
z
>
I 1. INVOLVE GENETIC CHANGE
0
w AMES TEST - POINT MUTATIONS
>
R DUBOW TEST - MOBILE GENETIC ELEMENTS
0
2. DUBOW TEST - INDUCTION OF BACTERIOPHAGE Mu
0
> IN E. COLI
z TWO WAYS
(A) THE TRAMSDUCT TEST - INDUCTION OF Mu
PROPHAGE TRANSPOSITION
W FUSION OF MINI Mu TRANSPOSE A GENE TO THE LAC Z GENE
(MORE SENSITIVE) (EtICODING P-GALACTOSIDASE)
3. STUDY WITH CIGARETTE SMOKE CONDENSATE
SURPRISE! REPRESSION OF Mu TRAtISPOSASE
4. UNDERSTAODING OF PlIE40P.E1,1100 IMPORTANT IN MUTAGEIIC
ACTIVITY OF SMOKE.
W018601

to
DUBOW PROPOSALS FUTURE STUDIES
z
z
t 1, CONSTRUCTION OF BIOSENSORS FOR DETECTION OF TOXIC AGENTS.
>
-i
0 NETICALLY PROGRAMMED
w 2. CELL RESPONSE TO TOXIC AGENT GE
> SELECTED GENES.
n INVOLVES TURNING ON OR OFF OF
n
0
3. MEASUREMENT OF RESPONSE
0 USE OF TH5-LAC TMNSPOSON TO INTEGRATE
LAC Z GENE OF E. COLI (BLUE COLOUR) OR
z LUCIFERASE, GENES OF VIBRIO HARVEY11 (LIGHT)
THAT
4, PREPARE LIBRARY OF E. COLI CELLS (PATENTABLE
RESPOND TO SELECTIVE TOXIC AGENTS.
5. LAC Z FUSION IN HUMAN CELLS POSSIBLE.
6, STUDY OF BIOLOGICALLY RELEVANT EFFECTS OF TOBACCO SMOKE-
WOL9601

IMPACT ON TIE BUSINESS
1. PLACES THE COMPANY IN A POSITION TO ACT IN AN
OBJECTIVE AND RESPONSIBLE MANNER WITHIN A CHANGING
SMOKING AND HEALTH ENVIRONMENT.
2. PROVIDE ASSURANCE THAT THE BIOLOGICAL ACTIVITY OF
PRODUCTS IS NOT INCREASED AS A CONSEQUENCE OF
UTILISING NEW PROCESSES, MATERIALS OR NOVEL DESIGNS.
3. PROVIDE A SOUND KNOWLEDGE BASE FOR THE FORMULATION
OF COMPANY POLICY ON SMOKING AND HEALTH.
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BAT Co LTD I TOBACCO LITIGATION