Details study including methods between the irritative quality of the by-products of identical tobaccos when treated with diethylene glycol versus glycerine. Tests done on rabbits. Examined the effects on the living tissue of the respiratory tract (the epithelial lining of this tract), the tissues of the nose, the nasopharynx, the larynx, hypopharynx, trachea, carina, right main bronchus, left main bronchus, lung. Includes lab tests performed on 6 cigarettes (Old Gold, Camels, Philip Morris, Chesterfields, Virginia Rounds, Lucky Strikes) to test whether they contained diethylene glycol or glycerol. Includes an appendix with microphotographs of experimental findings and normal controls. Concludes that there is more marked evidence of irritation effects due to cigarettes containing glycerine than diethylene glycol.
Report on In'vestigation
IRRITAN'T EFFEC'T'SI OF CIGARETTE SMOKE AS INiF'LUENCED BY THE
HYGROSCOPIC AGENTS DIET'HYLENE'GLYCOL AND GLYCERINE
SAMUEI, J' .: KpPET. zKY, M D.:, F'. A. C S.
Professor and Director of'the Department of'Otolaryngol-
ogy, New York Polyclinic Medical School and Hospital
A perusal of the literature on the irritant proper-
ties of cigarette smoke as influenced by hygroscopic
agents shows that s!ome work has been done from clinical
and pharmacological angleso Mulinos & Osborne (1)' (2) and
Wa11'ace, Reinhardt & Osborne (3), using, the conjunctival
sac of the rabbit as a test membrane and the resultant
edema produced as a.criterion, reported that cigarettes
made with dietlaylene glycol were less irritating,than those
made with glycerine. Hlaag (4) using the same method, re-
ported no d'ifference. Flinn (5) (6) reported a series of
clinical investi'gations and concluded that on smokers there
was a distinct difference in irritant effect of cigarettes
treated with dicthylene g].ycol and glycerine -- the diethy-
lene glycol cigarettes bieing definitely less irritating
(1) Pro:c. Soc. Exp., Bioll.& Med'., 1934, 32, 241-245
(2) N.Y. State Jour. Med., 1935',11,590
(3) Arch. Otolaryngolog3', 193Fi, 23, 3', 306-309'
(4) Jour. Lab. & Clin. Med. Jian. 19'37,22',4, 341-346'
(5) Laryngo'scope, 1935,45 149-154!
(6) Laryngoscopey 1937
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Ballenger & Johnson (7) and Ballenger (8), using clinical
observations, failed to find any di~ff erence. Carlson
(9),using salivation by smokers a~s a measure, came to the same:
conclusion as Ballenger & Johnson and Ballenger.
Without discussing in detail these papers, it is
very evident that the results these authors present leaves
the question open, because the results are controversial,,
In an endeavour to clarify the questions involved I make
a study ofthe subject from an entirely different angle
and one which had not heretofore been undertaken,
It is known that among the combustion products of
glyeerine are present glyceric, tartronic, oxalic,
tartaric, acetic and formic acids, together with acrolein,
Fromidiethylene glycol, among,the combustion products,
are foundiglycolic, oxalic and formic acids, but no
acrolein. It can be readily seen from the structural
formulae of glycerine and diethylene glycol that the
former, being a three-carbon, straight chain, trihydroxy
alcohol, could produce aicrolein, whereas from diethylene
glycoly having a chain made up of twoica~rbonss oxygen,
and two carbons, acrolein would not be expected. Acrolein
itself is exceedingly irritating and its presence could
(7) Arch. Otolaryngology, Jan. 193 7, 25, 1, 75-80
(8) Archo 01tolaryn'golo'gy, J'an:: 192 9, 219, 1, 115-123
(9) Proc o Soc. Exp. Biol e& Med,, 1937, 36, 302--307'
Page 3: goy81a00
be responsible for a differen.ce inithe irritant pro-
perties of cigarettes in whi.ch these two hygroscopic
materials were used, respectivelyo
Before any comparative study of'the irritant
effects of cigarette smoke as influenced by the use of
glycerine and diethylene glycol as hygroscopic agents
can be made, the various problems involvedimust be:
studied. First it is necessary, to_cancel out variables
other than the hygroscopic agents.
This is accomplish_ed..by using the same blend of
tobaccos made up into identical cigarettes., the only
difference being that in.one group the tobacco wasd treated withiglycerine and the other with
glycol -- both in the amounts regularly used on!pppular
ciigare ttes .
Observation of the habits of smokers shows that
some take a short puff and others a, long one, that some
puff'slowly while others take~ twice as many puffs pex~
unit time. The type of smoking also has an influence:
on th~e combustioniproducts and consequently upon!their
effects. To obviate these variables, a definite smoking
rate must be used whenitesting effects inr ordier to make
a; fair comparison withibut one variable.
Most important, however, is the choice of thee
membrane or tissue, the reaction of which is to serve as
a criterion for estimation of the irritant effects ofthe s,moke.-
In the study herein detailed', the effects observed
were the results of smoking the cigarette as a whole,
and noting determinable comparative histopathological
effects on the living tissue of the respiratory tract.
The two types of cigarette tobacco contents were tested,
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and the results compared, the tests being conductedd
under exactly alike conditions and undler controls,
established by exactly alike normal tissueso
Fizrthermore,, it was deemed necessary for the
accuracy of'the findi~ngs to make a fundamental study
----------- of the effects of th~e two types of processed cigarette
tobacco smoke upon living, tissuie of the respiratory
traict,, using a virginal tissue.
A word rega~rding my use~of the termi"virginal
tissue'~ is necessary. By this term is meant tissue
never before subjected to tobacco smoke irritation, This
type was found in the experimental animals selected,
Their tissues furnished material for autopsy findings,
not only of gross pathology, but permitted microscopic
histopathological studies to be undertaken. The
specimens are preserved for future study and demonstra-
tion. The sections were photographed as well as were
the microscopic~slides. The copies are part of this
Thus th~ere is at hand concrete evidence of definite
paithologicail reactions in the tissues examined. This
concrete evidence presented by the tissue themselves of
the body reactions to the two types, of processed cigar-
ette smoke are immeasurably more conclusive in making
determinations than are observations upon the human
nose, throat, pharnyx and larynx by a clinician. The
latter sees; and tells what he sees, but he cannot make
whalt he sees a matter of permanent record. In the study
detailed herein, the record is permanent and any compet--
ent microscopist can study the cuit slides and see thee
findings for himself.
Furthermore, one could assume~after careful exam-
iniation that the selected animal's respiratory tract was
Page 5: goy81a00
normal. Such an animal when killed furnished both gross
and microscopic evidence that it was indeed a,normal
Onlthe part of' humanibeings living in a civilized
community, a normal respiratory tract would be difficult
to find. A clinician examining a person can only state
that no pathology is found at his examinationio
Almost all human adults before they reach adult life
halve:had'numerous "inciidents" happen to them which leave
marks upon the respiratory tract.~ In addition it must
be remembered that most adult humans, in this time and
age, smoke tobacco in some formo Therefore, observations
made upon th~eir respiratory tissues a~s to the effects
produced by cigarette smoke must take into: accoun~t, those
reactions of a protective character which the bo~dy builds
to protect itself from irritating substanceso Th~e effects
of'smoking cigarettes must therefore, in humans, be pro--
duced over and beyond the protective barriers which nature
already has established in the individual- Greater effects
observed in one individual human a~s compared to those
observed in another, may depend on the extent to which the
given individual has established'his own tissue defense
mechanismsa This statement must however not be construed
as a criticism of comparative clinical observations made
upon humansto record' their perceivediphyaical reactions
while under clinical study and control to test the effects
upon them of'cigarette tobacco smoking:
My study was concentrated upon the respiratorg tract,
in general, and upon the epith.elial lining of this tract
- ---- ----- - ---- in particular.,
This epithelium, and its ciliary covering especially,
is probably the most vitally important tissue of the body>
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In smoking,, especially when smokers inhal'e the
tobacco smoke, this vital tissue comes into direct contact
with the epithelium of the upper respiratory tract, andd
often with that of the lower parts of this tract alsol.
Thie rabbit was selected as the test animal~ This wais.
done because of its convenient size for handling, its
availability, and particularly because the epithelial
lining of the rabbit°s respiratory tract possesses exactly
similar features and histological cell characteristics to
those found in humanse The rabbit°s epitheli!an lining of
its respiratory tract, its ciilia:; its cell character-
istics, subepitheliial stroma are identical with those
found in Mumans~ Obviously the anatomical structure of
the rabbit differs from humans, but its mucous membrane
and component parts nevertheless are identical tissue.,.
As far as ascertainable, rabbits of'the same sex
(miales) equal size and weight, and even coTor or fur were
The animals were tracheotomizeda That is, a small
cannula was introduced into the wind-pipe (trachea) and
the tobacco to b~e tested was s!moked', by the animal using,
the automatic smoking machine devised by Dr.Arthur Walter
Proetz of St. Louris,
The tracheotomy tube in itself caused some local
irritation~ These!melchan,ical irritation factors being,
present in both series of cigarette tobaccos tested can--
celled each other out. This mechanical irritation factor
could easily be visually recognized. It was totally dis-
regarded in making determinations fromiboth th~e gross and'
the microscopic findings of the tissue reactions from
both series of cigarette tobacco contents which~were
Another factor needs notice, In the test animals
Page 7: goy81a00
the insertion of the tracheotomy tube doles not entirely
block,the upper respiratory tract as it does to humans
upon whom a tracheotomy tube is used to permit respira-
tion because of some tumor or other obstructionof res-
piration through the larynx and the nose or mouthe
Because the tracheotomy tube in these test animals
did not entirely block off the upper respiraltory area it
gave our t.est ain additional a~dvantage, The upper respir=-
atory tract is open andfree, in the test animals, and
the tested smoke can be exhaled through the nasal pass-
ages. This made possible the study of the effects of the
tested smoke, on, the tissues of the nose, the nasopharynx
and the larynx.
The automatic smoking ma~chine walst set to smoke "puffs"'
at the rate of'two puffs per minute. By "puff" is meant
an inhalation, an exhalation followed by a period of rest
The animals were permitte4to smoke for only one
hour eachiday in those tested for more thanone hour of
smoking ti'me o
Following each smoking period the animals were re-
leased in their cages, free to roam abouit, take their
natural nourishment by moutha They reacted well, andd
acted in no way different from other rabbits in the laib--
oratory animal quarters.
A few animals died of pneumonia.contracted inter-
currently during,the experimental work~ An autopsy was
performed on these. The resultan~t findings are includedi
in my report. Curiously all those that died were those
who had inhaled the gllycerine treated tobacco.
While this finding has no effect upon the conclusions
drawn in thle problem upon which I am here reporting, it
RABBITS STUDIED: During experiment, after
GROSS: Nlasal chambers
----- Pharynx and Hypopharynx
Right and Left main bronchus and'
MICROSC'QPIC': Sections taken from
Right and Left main: bronchus and
)Entire respiratory system,
Gross )Nasal Chamber
)Trachea, Carina and Bronchi
STUDY OF NORMAL TO PROCURE NORMAL SPECIMENS
Two Rabbits Used.
One rabbit killed' and entire.respiratory system
was preserved in formailin~
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Second Rabbit killed - respiratory system removed and
.sections made from the following,diesign-
ated areas for microscopic study:
Right main Bronchus, Bronchioles,
Covered with stratified squamous epithelium
Sub-epithelial layer containing blood
vessels, connective tissue an~d, cells,
occasional plasma cell, mucous gland ducts.
Numerous mucous glands~
S'quamious epithelium except in ventriculus
area, where ciliated mucous membrane with
numerous ga~blet cells are presento- Very
thin sub-epithelial layer - cartilage -
muscle fibre striatedi,
Pseudostratified ciliated column epithelium
with several goblet cells. Benealth this
layer is a thinicompact connective tissue
Sub-.epithelium consists of loose areolar
type connective tissue rich in blood vessels,
Connective tissue cellscartilage.
Simple ciliated columnar eplitheliumlin folds_
Some goblet cells.
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Sub-epithelial layer (loose areolar) contains
blood vessels, connective tissue cells, somee
smooth mus.cle fibres, lymphoid tissue, plasma
Right Main Bronchus: Bronchiolies - Lung_
Silmplc Low columnar ciliated epithelium on a
balsemenit mlembraneo Some goblet cell -
submucosa is a loose areoliar connective
tissue containing thin blood vessels, con-
nective tissue cellso (Some plasma cells)
mucous membrhine in folds posteriorly where
some smooth muscle fibres are present -
Very low simple non--eiliatedi epithelium
(Simple Siquamous) alround air spaces< Some
spaces contain geletinous material with aa
few polymorphs, and red bloodi cells in it.
In some areas around bronchioles may be
seen collections of'lymph cells making up
lymphoid patches:, In these areas maly bee
seen occasio al giant cells probably for
fighting off infectiono
Left Main Bronchus - Lung - B'ronchioles:
Description same as for right siide,
In which animal was smoked with Diethylene_gjycoj
treated cigarettes for one hour through a tracheotomy
tube, killed,, and studied.
Cigarettes u edicontained Diethylene-glycol Q
Respirations 34-36 per minute O
Smoke inspired through *racheotcrmy tube and expilred 0
Page 12: goy81a00
partially through nostrils and partiailly through!
No moisture was observed from nostrils.
Autopsy revealeri no gross changes
Hypopharynx: Somie hyperactivity of mucous
Larynx: Epithelium normal - very little
infili,,ra~tion of' sub-epithelial
layer Blood vessels slightly
Trachea: No changes
Carina: Some secretion in lumen
Right Main,Bronchuis: Narmail epithelium
Slight oedema of submulcosa
Slight perivascular infiltral-
EXPERIMENTAL ANIMAL #2'
Rabbit "2G" September 28, 1942.
Animal was smoked for one hour with cigarettes
in whichi tobacco contained glycerinic-.
R'espiration 34-36 per minuteo
Vapor was observed in inhalation tube (tube
connecting smoke reservoir to Trachea of
Showed slight discoloration of the
trachea with dilation of blood
vessels of the trachea and carina.
Hypopharynx: Hyperactivity of'the mucous
Larynx: Sopne oedema of the sub-epith-
Dilated blood vessels-
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Perivascular infiltration with
Trach'ea: Increase activity of goblet
cells. Some oedema of sub-
Cariina': Inerease in the goblet cell
Some engorgemen't of the blood
Mostly mononuclear ee11s.
Right Main Blronchus:
There is an increase in gobliet cell
Some polymoxphonuclear cells in the
Engorgement of'the blood vessels with
Left Main B'ron'chus ~
There is some interference with the
There are some golymorphonuclear cells
infilt~ating the mucous membrane.
Marked perivasula'r infiltration>.
Some polymorphonuclear cells as well
as connective tissue cells in the sub-
EXPERIMEN'TAL ANIMAL #'3'
Animal was smoked for three hours~ One
hour a time for three consecutive days.
use~d contain~ed ID'iethylene glycol.'.
Respiration. 5u' per minute (increased respir-
ation) Vapor was observed in the inhaling tube
after the fifth cigarette..
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No vapor was observed during the second and
third hours of smoking.
Autopsy: Revealed some dilated blood vessels in
the lower portion of the trachea. There wass
some small pinpoint hemorrhage in the lung~so
Hypopharynx: No changes in the epithelium.
Hyperactivity of the mucous -
gl andis .
Some engorgement of the blood.
in the sub-epithelial layer,
Larynx: Mucous membrane appears normal.
There is some secretion in the
lumen of the larynx.
Blood vessels dilated.
Very little perivasculair
Trachea: Ciliary activity diminished.
There are some polymorphonu-
clealr cells in the epitheliall
The sub-epitheliall layer shows
an occasional polymorphonuclear
cell close to epith~eliu,m.
There is nolengorgement of the
There is some oedema of the
sub-mucosa layer; most of th~ee
cells are plasma and connect-
ive tissue cells.
Cairinai: There is very little inter=
ference in the cilia of'the
The submucosal shows very
little cell infiltration.
There is some engorgement o~f
the blood vessels.
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Right Main Bronchus: There is some secretion in
There are a few polymorphonu-
clear cells in the epithelium.
The submucosal reaction is
very mild, showing very little
Left Main Blronchus': There is some interference
withicilia of the mucous
There is some engorgement of'
the bloodi vessels.
_ Very little connective tissue
reaction and perivascular
Animal was smoked for three hours. One
hour at a time for three consecutive days. Cigarettes
Respiration: 34-36 per minute.
Autopsy: Revealed some grayish areas over the
mucous membranie of the trachea.
Mi¢roscopicFi,ndings:Hy,popharynx: No change in thie epithelium.
Hyporac tivi ty o f the mucous glands.
Some engorgement of'the blood',
vessels in the sub-epithelial
Larynx: Interference with ciliary activity.
There is an increase in perivascular ).ainfiltration; more marked than in Q,
,, 38,; o 01
Trachea: There is some uliceration of the Q
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Submucosa shows increased oedema,
numerous polymorphonuclear cells
and' periva~seular infiltration.
Carina: Loss of cilliary activity, Mucous
membrane denuded in areas. Sub-
mucosa infiltrated many pol'yso
Right gain Bronchus: The mucous membrane and submucosal
layer are invaded with polymor--
Increased connective tissue react-
Left Main Bronchus Interference with cilia~ry a~c.tivi~ty~
. Epithelium infiltrated with poly-
Perivascu2lar and submucosal
(this shows evidence of infectioni,)
EXPERIMENTAL ANIIiMAL #5
Animal was smoked for six hours. One hour
at a time for six consecutive d'ays. Cigarettes contained
Respiration: 3!6-40' per minute.
Small amount of vapor was.observed
inithe inhalation tube (tube con-
necting smoke reservoir to Trachea
No vapor or moisture after the
first hour of smoking> During
the fifth and sixth hours of
smoking respiration increased to
55' to 60,per minute.,
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Autopsy: Showed a little mucous in the Trachea.
Some dilated blood vessels in the
larynx. A few hemorrhagic spots in!
Hypophalrynx: Epithelium appears normal.
Some increased mu~eous gland
Blood vessels show engorgement.
Very little sub-epithelial
Larynx: Pus in th~e luimen.
Very little interference with
Slight sub-epithelial tissue
Trachea: Increased activity of the goblet
Some loss of'ciliary activity.
Some (jedema of the submucosa.
Carina: Diminished ciliary activity.
Increased activity of the goblet
Very little infiltration of the
Right Main Bronchus: Interference with ciliary activity.A few polymorphonuclear cells
infiltrating the mucosa.
Connective tissue reaction is
Left Main Bronchus: Some secretion in the lumen.
No infiltration of the mucosa.
Connective tissue and peri-
vasculair reaction slight~
Page 18: goy81a00
EXPERIMEN!TAL ANIMAL #6
Animal wa's smoked'for six hours. One hour
at a time for six consecutive daiys~ Oigarettes contained
R'espiration: 45 - per minute,
During the third hour of smoking,,tobacco
stain was observed around' the nostrilsA
millky secretion fro,m -the lieft eye wa's also
This same observation' was noted during, the
fourth',, fif'th and sixth hours of smoking,
Respirs.tions increased to between 50 and 688
times per minute dhiring, the fif'th and sixth
hours of smoki~ngo
Autopsy: Showed a large amount of mucous iin the
trachea and lungs The blood'vessels of the
trachea were markedly dilated, The sku11
was dissected so as to bring into view the
interior of the nasal chamberso These
cavities were lined with a thick mucous
film (pyogenic membrane, ) This membrane
was attached to the mucosa of the nose on
th'e l e f t s lide .
Hypopharynx: Pus collection in the lumen0
Mucous gliands hyp'eracttive:
D'ilated blood vessels in the
Larynx: Phs in the lumen:
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Sub-epi thelial layer shows
infiltration of all kinds of
cells including polymorphonuclearo
Loss of ciliao
Mucous membrane ulcerated in areas.
Blood vessels of the submucosa dilated.
Carina,: Pus in the lumen.
Loss of'ci'~liary activity.
Engorgement of vessels in submucosa.
Increased perivascular infiltration.
Right Main B'ronchus: Muchpus in the lumen.
Loss of cilia.
Mucosa ulcerated iniareas.
Mucosa and submucosa inifiltratedd
Left Main Bronchus:
Polymorphonuclear cells infiltrating
Submucosa shows muchiless reaction
than th~~eright main bro~nchus:o,
EXPERIMENTAL ANIMAL #7
Animal was smoked for six--hours~ One
hour at a time for six consecutive days. Cigarettes
contained diethyleno glycol.
Respiration: 40 per minute,.
Particular care was taken to notice any tobacco
stain around the nostrils~ Only a slight brownish
discoloration of th~e mucous was found-
Autopsy: Only a slight amount of the congestion of
the mucous membrane! of'the trachea was observed.
The skull was dissected so as to bring into view,
the interior of'the nasal chambers. A little
mucous was.present but no pyogenic membrane could
be found. This specimen was preserved in formalin
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Respiration~ 32-36 pier minute
EXPERIMENTAL ANIMAL #8
Rabb i t "86"'
Animal was smokedifor one hour, withitobacco
EXPERIMENTAL ANIMAL #9
to become part of the gross exhibito
Rabbit died - before second hour of smoking
could bie startede
Autopsy: Reveailed oledema of both lungs and
a large amount of muco-pus in the
This specimen was fixed in formalino
Animal was smoked for one hour,, with tobacco
Respiration: 32-36 per minutea
Rabbit died the following morning.,-
Autopsy: Revealed acute pneumonia.
EXPERIMENTAL ANIMAII, #10
-------- ------ ---
Animal was smoked for four hours, one hour
at a time for four consecutive days withi tobacco: contained
Respiration: 22-40 per minutea
During the fourth hour of smoking - after
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the third hour of smoking, tobacco stains
were noticed around both nostrils. This
rabbit died before the fifth hour of
smoking could be started.
Autopsy: Nasal passages contained a fair amount of
muco-pus. The trachea showed marked in-
flammat,io.n and a large quantity of muco-
pus.- Both lungs were conge ted. Muco-
pus was present in the bronchi. Consolida-
ted areas in the lung gave evidence of
Microscopic: Sect,io.n from th~e right main bronchus and
surrounding lung tissue revealed much
purule,nt secretion in the lumen of the
bronchus. The mucosa is denuded. Sub-
mucosa markedly infiltrated with poly-
morphonuiclear cells. Lung,tissue shows
numerous consolidated areas. Evidence of
acute infectious pneumonia.e
EXPERIMEN'TAL ANIMAL #11
Rabbi t 1'111G"'
Animal was smokedifor two hours - one hovi!r
at atime for two consecutive days with,tobacco contained'
Respiration,: 44 pier minute.
Moisture from bothinostrils were noted,,
andian increase in salivation. Tobacco
stains over the nostrils during thesecond hour of smoking. This rabbit diedd
before the third hour of smoking could
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Autopsy: Revealed large amount of'muco-pus in
trachea and bronchIe-
Lungs showed evidence of'pneumonia.
EXPERIMENTAL ANIMAL #12
R'abbi t "'12G"
Animal was smoked for three hours, one hour
at a,time for three consecutive days withicigarette
tobacco contained glycerine.
Respiration: 34-50 pe-rminute.
This rabbit diedibefore the fourth hour of
smoking could take placeo.
Auitopsy: Revealed trachea and bronchi filled
with muco-pus. Lungs showed evidence of
(#8 - 9 - 10 - 11 and 12 were placed in
formalin and preserved for gross exh~ibit.)
Rabbi t "'13D I"
This animal was smoked for three hours, one
houi!r at a time for three consecutive days using, cigarettee
tobacco contained diethylene glyc,ol,
Respiration: 20 -40 per minute,Rabbit showed no evidence of discomfort.
No moisture formed in smoking tube. No
increased saiIiivaition: Rabbit wais kililed
a~fter the third hour of' smorking,.
Autopsy: Revealed nolgross changes in the
respiratory tract. Specimen preserved for
Page 23: goy81a00
---- -- ----- ------ ---
Animal was smoked for six hours, one hour
at a time for six consecuitive days using cigarette
tobacco contained! giycerine.
Respiration: 20-28 pier minute.
During second hour of smoking, left eyee
showed a, miIky secretion. Increase mucous,
secretion from nostrils. During third
smoking hour, mucous accumulated in thie
trachea, and was removed'with a suction
tube,- Sub-maxillary region a little
swollen. Milky-secretion from the 1ef;t
eye, and small amount of tobacco stain
around the nostrils.- During fouirthy
fifthiand six,th hours of smoking, the same
findings as above were observed. Tobacco
stains around the nostrils formed after
Rabbit was killed after the sixth hour of'
Autopsy: Revealed muco-pus in the tracheai and
bronchi and nasal passages.
This specimen was used for photog,raphy.
EXPERIMENTAL ANIMAL #15
Animal was smoked for three hours. One
hour at.a time for three consecutive days. Cigarettes
Respiration: 35-50 per minute.
Autopsy: Revealed some mucous secretionlin
Page 24: goy81a00
nasal meatio, Numerous punctate areas over
lower part of the trachea coaIescing into a
pyogenic membrane. Some punctate areas over
mucous membrane of right and left main
Mucous membrane of trachea and bronchi appear-
Rough Test of the Cigarettes on the Public Markets
The two series of'tobaccos tested and upon which
I reported above were delivered to the laboratory by
the Philip Morris Company.
To test the cigarette tobacco in the cigarettess
as they are sold upon.the open market,.I bought six
different brands of cigarettes on the op~en market in
September, 1942. I emptied their tobacco contents and
assigned a code number to each, as follows: -
#1~ Old Gold
I sent the tobaccos thus marked to, Dr. Samuel
Ro~senzweig, of the Synochem Corporation, 80 Fifth
Avenue:,, New York C'ity. I happen to know this research
chemist very well, andhave full confidence inihis
intellectual integrity- He was unaware of thie names
His report is as follows: -
Page 25: goy81a00
'S T A T E M EI N T^"
concerning the resu!lts of the qualitative analysis of
six samples (#1-#6) of tobacco
provided by Profo Kopetzky.
"After a study of pertinent literature, ten gram
samples were extracted with acetone following the pro-
cedure of chapman, AnaSyst 51, 382 (1926). In quanti-
tative work, the final extract obtained upon evaporation
of'the acetone and remova'l of resins should be subjected
to a Zeisel determination,"
"However, at this poinit, it was decided to estab-
Iiish rather the chemical nature of the moistening agent
tha.n its quantitative amount. Therefore, the residues
were used' to prepare - naph'thyl urethanes following the
procedure of B'ickel and French, Journ. Am Ghemo Soc, 48,.
747' (1926) . "
"'In each case,, 05 g',., samples of the extract were
mixed in th'e cold with' 0, 5 g of - naphthylisocyanate,
and the crude urethanes were carefully recrystallized
"'From the observed melting points, it is concluded'
that the samples #1 and #3'contained diethylene-glycol,
whilie the samples #2, 4, 5, &6contained' g~ycerol. If'
the observed'' amounts of urethane should permit to d'raw
additional conclusions, it would app'ear that sample #3
contained more diethylene glycol than sample #1, and
that samples #5 and #6 contained'more glycerol than
samples #'2 and #4
Note:--Samplie #3 was the tobacco co tents which I
removed from a carton' of Philip Morris cigarettes,
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connective tissue cells, (c) blood vessels, and (d)
Five animals diied'before the experiments could be
completed. In all these cases cigarettes moistened
withiglyeerine were used. In all these cases acute
tracheai- bronchiolitis and pneumonia were the cause
FINAL OBSERVATION AND CbNCLUiSIO'N
From, this study o-f"the irritation reactions from
exactly alike cigarette tobacco manufactured, on the
one hand using,glycerine, and on the other using
diethylene glycol, and noting, the effects produced upon
the living tissue of the respiraltory tract of rabbits,
it is very evident that more marked evidence of irrita-
tion effects are present in these tissues when subjeeted
to the smoke of cigarettes containing glycerine than
from those containing diethyl,ene glycol,
Finally, the data presented in this paper should
tend to end any controversy in regard to the effects of
hygroscopic agents in cigarettes on the..irritant pro-
perties of' smoke. The method of manufacture has a,
distinct and measurable effect on the irritant properties
of' smoke I C'iiga~rettes in which diethylene glycol is used
as the hygroscopic agent in the above study have beenn
shown to be less irritating, than those in which glycerinee
Grateful acknowledgement is made to the Laboratory
Department of the New york Polyclinic Medical Slchool
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and Hospital for extending~, to ~ my staff and to me~~ its
Likewise I aim indebted to Dr. Julius Bell,
Instructor in Embryology and Histology at the above
named institution for making the sections' and controll-
ing,their preparation for photography, and for his
active cooperation and interest in checking,upon both,
the gross and the microscopic findingsa To Mr, Joseph
D. Ha~ckel of the New York Medical Film Guide I am
indebted for his time consuming`Work inisetting the
microscopic specimens in permanent visual records. His
phoitography has made easier the preparationiof this
report-by producing a faithful record, by untouiched-up
photography, of both the histoliogy and the pathology
exhibited.in the specimens.
SAMUEL J. KOPETZKY, M.D., F'.A.C,S,
April 2, 1943
71 East 80 Street
New York City
- 2'8 -
N 1 Upper Respiratory tract. 4X - Normal showing
anterior (Anferior) turbinate, middle tur-
binate, tongue, nasopharynx, oropharynx..
'N! 2' Respiratory Tract. (lower) 4X. - Normal,
Epi'gDottis, Larynx, Trachea, Carina, Right
NainiBronchus and Left Main Bronchus and
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N ' 3 Trachea. (18!0X),JMIncosa1, ciliatory columnar
epithelium with goblet ce11s. - Submucosa
and cartilage and a blood vessel.
Ni 4 Trachea - higher magnification 8010 X.
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N 5 Bronchus and Lung, 18'0 X- C'iliated columnar
epithel'ium with goblet cells, with thin sub-
mucosa'li layer, containing some smooth muscle
fibres, cartilage and lung tissue and blood'
v essel (branch of pulmonary artery) wi'th
1D 1 Tlrachea' - ti8OX - After one hour of smokin!g
Di'ethylene Olycol treated tobacco.
Submucosa slig'ht oedema
6light amount of ti'ssue cel'1 reaction.
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1D 2 Bronchus& Lung - 180 X - After one hour
of smoking IDilethylene Glycol treated tobacco.
No pathological changes.
3D 6 Some interference withicil!iary activity of
mucosa, few polymorphonuclear cells in
mucosa. Slight submucosal and lung tissue
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5D 1! Trachea - 180 X - shows some interference
with ciliary actYvity and some infiltration
of polymorphonuiclear cells. Suibmucosaa
slightly oedemaltous - increasedi cel!llular
activity andivascular engorgement. Slight
amount of perivascuilar infiltration.
5D 2 Higher Magnif'ication. BOO X - finding the
same as 5D 1.
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5D 3 Bronchus and Lunig~ 180' X'.
Diminished ciliary activity and infiltration
of'mucosa with a few polynuclear celils,.
submucosa shows sli~ght connective tissue
reactions with increasedl perilvascular
infiltrations. Slight lung reaction.
7D 1 Respiratory tract.
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7D 2 No. gross pathology.
703 3 Trachea and Lunig - No gross pathology, a
slight congestion appears in the mucous
membrane of the tracheal.
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1i3D 2 Nio gross pathology 4X.
130 D 3 No gross paithollogy 4'X. .
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13D 4 Trachea 180-X ..
Very slight goblet cell activity.
Slight increasen in oedema and
13D 5 Trachea 800 X - ANagn!irien same as 13D 1.
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2G 1 Tra'chea 180 X- After one hour of 6lycerine
Increased'gotilet ce11 activity in mucosa
Oedema of suhmucosa.
Marked increase in cellular activity.
Increased' cel!lular activity..
.~ . . -
2G 2 Trachea - Hig,h Power of' Same as 2G 1
slame area, same find3ngs.
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2G 3 8'ronchus & Lung - 18OX - Increased goblet
ce1i1 activity an(l' mucous secretiloni. Sub-
mucosa shows increased lymphoid cells and
connective tissue cells. No change in
IG 1' Trachea. Lonw Power. 7'2X - 3 hours Glycerine
treated Tobacco - Purulent secretion in
luimen of trachea, - mulcosa shows interference
with celiac erosion aind ulceration.
Submucosa shows marked oedema infiltrati'oni
with connective tissue andipolymorphio.nuclear
cells - Di1'ated,blood vessels andiperi-
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4G 2 TrachW18D X - Three hours - Glycerinee
treated Tobacco - finding same as 4G' 1.
4G 3 Lung - 1&0 X Three: hours - Glycerine
Purulent secretion in"bronchial lumen.
Interf erence with ciliary activity mucosa
ulcerated in areas marked infiltration of',
submucosa with"P. N. M. celiLs, marked
perivastular and lung tissue infil!tration.
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6G 1' Upper Respiratory Tract. 8 X - After 6 hoursg smoking withiGlycerine treated Tobacco,
showing Pyogenic membrane over inferior
turbinate extending into inferior and
middle nasal meati.
6iG 2 Trachea - 180 X'- After 6 hours.smoking
with Glycerine treatediTobacco showing
Pyogenic membrane attached to mucous
membrane, loss of ciliary activity.
Ulcerated in, areas markedd iinf i ltrat ion
of submucosa vascular diilatation and
marked infiltration with numerous GcuDy-
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6GI 3 Blronchus and Lung - 180, X. After smoking
with Glycerine treated Tobacco, showing loss
of ci~Liary activity, mucousme,mbrane ulcerated
in areas,, pus in bronchial lumen, increased
perivascular and lung tissue reactions.
Engorgement of pulmonary bloodl vessels..
6G 4 Trachea - 8!010 X - After six hours of smoking
with Glycerine treated Tobacco, showing
pyogenic membrane attached to the mucous
membrane, loss of ciliary activity, ulcerated
in aireas marked infiltration of the sutr-
mucosa, vascular dilitation, and markedisub-
mucosali infiltrate with numerous poly-
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15 GI 2 lDpper Respiratory Tract 4 X - After 3 hiours
G1'ycer.ine treated Tobacco - several smai11
punctate areas on mucous membrane of
milddle meatus and inferior turbi,nate.
15G 3Trachea and' Lung 4 X - After 3 hours
Glycerine treated Tobacco - numerous
small ulceraltilons ever lower part of
trachea coalescing to and covered with
a, pyogenic: membrane.
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14a The gross specimen (14GI) after six hours
of smoking-°with Glycerine treated Tobacco
showed! large areas of uilcerati'on of' the
treachea covered'with a thick py ogenic_
14 G Lung and Trachea 4 X - Large ulcerated area
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14'G Head - After 6 hours with Gllycerine treated
'Tobaccos showed: Purulent secretions inn
the anterior (inferior) meatus and over
the inferior turbinate.