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REPORTNo. 8-6 (Final Report)

CONTRACT No- DA - 92 - 557 - FEC -39659

3rJDIES OF ESFIRATORY DISEALF3 DUE TO AIR POLUlTIONsi W. TOMcLO-YOKOHAMA AREAb y

ob y

Tatwaii Ishizaki, M.D.

lacturer of Clinical Allergy

University of Tokyo

School of Medicine

Department of Physical Therapy

& Medicine

D cemer 1966

U. S. ARMI RESEARCH AND DEVELOPMENT GROUP AnFAR EAST

APO San Fracisco 96343

BEST AVAILABLE COPY

REPORT No. J-i88-6 (Final Report)

C0I1A'CT No. DA-92-557-FEC-3C659

DA Project/rask Arcoafiork Unit No. 2110l14SlEB71D 00 04~3FE

RESPIRATCRY DISWS~ D"1 TO ADl~ POLLUTION IN

TOKYO-YOKOHAMA ARZA

by

Tatsuabi Inhisakig , D.lecturer of Clinical AllerF7University of TokyoSchool of MedicineDopartnt of PbysioalTherapy & Miedicine

Dooomber, 1966

t,1. S. AWX RSIACH A!M MVZLCPWINT MOUPPAR SAME

APO San Fra:r!isoo 9634~3

Distribution of this document is unlinited,

C 0 N T E N T S

4.

* PA'T I The inf luence of 3u]_fur dioxide to ath.mt.cpae-nts measuved by the r a:.1 o1 f iriye .a .n • . . . . . . . . . . . . . . . . 2lMterials and Methods ... .. .. .. .. .. .. . . . 2

* Revv,.ts . . . . . . . . . . . . . . . . . . . . . . . . 4b ~ . .s o . . . . . . . . . . . . . .. . . . ..Z,- - r,iid Ref,,rcrcer .. . . . . . . . . .6 . . . . . . . . 6

.. l. ... . . . . . . . . . . . ..; .i .. . . . . . . . . . . . .I . . . . . . . .

.... . .I .. . . . . . . . . . . . . . . . . . .. .ir

.. . .~ .. . . . . . . . . . . . . . . . . . . .. .Fi . . . . . . . . . . . . . . . . . .* . . . . . . . . .3.

TabIe 2 . . . . . . . . . . . . . . . . . . . . . . . . . OIi" ble 3 . . . . . . . . . . . . . . . . . . . . . . . . . . . .15

* S .. . . ................. :1W

RJT? II Changes in mehanical properties of ,.Liea pifin exm.rimental as'hma induced by acctylcholineor histarrine .... ......... .... . .i7

.ehods . ... . ... .. ...... ........ . 17? ,s and Discussion .... ..... ........ .. .1

. .,,. . ........................... 1

............. .............. 20. .. .. ....................... 2

". . " .................. .... .. .. . . .2 5

vA, II influenc! of sulfu: iiox'de expov':wo toexreriental asthma in guira .igs . . ..... .64n!... . .. .ehods....................26

Ri#su' s' "nd Dis-'-,ssjoie. ..... .................. 27

. . .. . . . .. . .

Abstract

Part I.Fifteen normal subjects aid eighteen asthmatic patients were tested by spiro-

graphy and body plethysmography, and changes of' airway resistance oeforo and afterinhalation of sulfur dioxide in three and six minutes were followed and anmlyscd.

The mean of variation of airway resistance was 1.85 in normal subjects and2.03 in asthmatic patients, and S.D. of variation of that was i0.2 in formar and-1.67 in latter. Airway resistance was higher in moderate or severe case: of di-sease than in mild case3.

Asthmatic patients showed increased sensitivity to sulfur dioxide at 5, 13, 20ppm in the air during the period of three and six minutes inhalation wile no changewas recognized at 13 ppm in the group of normal subjects. A correlation betweendaily variation of airway resistance and daily changes of the atmospheric contentof sulfur dioxide was noted in a asthmatic subject.

Part II.In experimental asthma, change of mechanical properties ef the lung of guinea

pigs was studied in relation to dose of administered histamine and acetylcholino.To-Lai flow resistance and pulmonary resistance increased abruptly at the time ofshock as much as twice of the control value, while pulmonary compliance began todecrease on in)'lation with the concentration around one sixteenth of shock con-centration and thereafter almost linear relationship was found between pulmonarycompliance and logarythm of histamine or acetylcholine concantiation.

According to those results, the influence of sulfur dioxide exposurc to ex-perimental asthma was studied in guinea pigs, measuring bronchial sensitivity tohistamine by the change of total respiratory resistance. However, no significantchange of bronchial sensitivity to histamine was observed among the groips exposed.The results give us an impression that the air-pollution mieht be one of the aggra-vating factors of bronchial asthma but have little role in the etioloa.

' I

STUDIE CP RESPIRATORY DISEASES DUE TO AIR POLLUTION IN THE TOKYO-YOKOHAMA AREA

It has been found in the previous reportsl)2)that the symptoms of bronchialasthma and chronic bronchitis increase with higher concentration of juspended par-ticles and sulfur dioxide in the air, and moreover, an impression was given thatthe inhalation of sulf'u dioxide increased bronchial sensitivity to acetylcholinein guinea pigs in which changes in mechanical properties of the lung such as lungcompliance and airway resistance were measured.

In an attempt to explore the above mentioned results, further investigutionhas been aimed to clarify, whether airway resistance of asthmatic patients is in-fluenced by the presence of air pollutants in Tokyo (1), and whether the mechanicalchanges in the lung in guinea pigs induced by histamine inhalation are influencedby the inhalation of noxious agent as sulfur dioxide (2).

PART I. The Influence of Sulf~ur Dioxide to Asthmatic Patientsmeasured by the Change of Airway Resistance

Pulmonary obstractive disturbance observed on asthmatic patients was measuredby spirography and body-plethysmography as well. However, airway resistancemeasured by body-plethysmography was mainly discussed in this part and also dis-cussed in relation to other parameters measured by spirography.

Materials and Methods

1. SubjectsFifteen normal subjects and eighteen asthmatic patients were tested by

spirography and body plethysmography. Normal subjects were healthy labcratorypersonels having no respiratory disorder due to specific or non-specific diseases.The asthmatic patients were chosen from the Outpatient Clinic of the Department ofPhysical Therapy and Medicine, University of Tokyo, School of Medicine. Their sexand aga distribution were listed in Table 1 and 2.

2. Methods1) Spirography was performed using Benedict-Roth Respirocater which capacity was13-5L and it's recording speed was 44mm/beo. Measured parameters were vital capa-city (vC), a percentage of vital capacity to the predicted volume (%VC), forcedexpiratory vital capacity (FEV), ore second volume of WE ( 1'i.) and a percentageof EV1.o to the .FV (FRVi.O%

2) Airway resistance was neasurad by e body-pletoysmrai ph with Nochisukis methodwhich was a reformation of DuBois' or44.inal method. Applied principle was basedon the theory that airway,q tnco (R) in e ati of' as-irway pressure (P)durin flow to airflow (V). ) Measurinc -woodure is stated below.

A subject sits and breaths inside an airtight box, in which pressure of thebox is the mirror image of alvelolar presure. If the ratio of alveolar pressureto box-pressure is laown, airway resiatance will .oe calculated by aeasuring airflowand box-pressure. Airflow can be masured directly by asie otachopmph.

2

T11 D'IIB;z LB ' air---- o.. p-tuo*ta-.ho6r-nh is shut~ during panting,so that J,.vrolar vrizecan bc :zelat-ed t. thrf plethysmographic. pressure becauseof 11o tir w -at I ~;th - r.: , r. Vi-i rt,',*. ', in YoellizukVLb modified method,an art-A..icie2 '.esint:-rc , &'w!iui aJ1~p i. . know.n, ii inserted 'cetwpen the mouth-piece ~a the ,uo 6~~ .zr-int; L,;i. A, :-.ingly, resultant phacediffei-enue bet-w r _- .W4~~1,4-i pr> . -z, i.e. siY:.'nola: pressure, and airflowis related to the pnar~a 6-fI-,er.e cct.raa ~2a n ~riiilresistance is re-moved.* Alve 112 tan plt; ,.-ia ~he~ c~st' e e~irulated fron such para-meters as ;Ihfisi P_'-_.I1-7, a±;.1wit pressure.

The. ivan.!e ap-z i. zsho v 2.. ?A * A fid points -In t e apparatus wasas follows.(1) Anetf~s1 's~~ ran i~ioecte. 'b7?-...2n the mouth-;.I--fe snd the pneumota-

chorap. cK'.i!:rt.- ~ontroilei by r'L crerat.,,r fromn outezde..(2) Aszi.a t Ar--,J.bt box --as pl-Lcel i!,. fr ,nt o: the face of a subjects sitting in

the bn'x. Ai.r i4z1 thfl~t~ box was,! connectefr, to that of plethysmograph througha tuin of 5orn 9-id 103c ~]O i ini le: -tnL, aa~d temperatur of' it was kepta' ?70 vth vr-vDcn..: !atu..'tici,. Tnta cwr.ll b-:;! vgas designied to avoid a devia-tion o; meaur.caent.' tv# -*- the _- 2Lrn-. of tu) eraturpe and humildity betweenexhaled air and air iii the bo. of' pleLsm.)-aph.

(3) Meast.roment ox' aj.rwqy res4.stancP by 1ochiv.ukils method was done in following;rinciple. A oinbject cat and waz; askel to part thz'on~h the pin-eumtachographin the body-plet.grzcmograph. A-rIz and th., bsx-prpscurr were recordedsimultanecufjy -i. rec'ordnq paper trrov,-,n tbe ptric. in which artificial re-sistance was i.zecrtod t'.4te=i mre ne:-*,oca an.1 thi -!:.eurnotachograph and re-maoved dUring -the fast ehal11w breathing.

Measuremnent was carriLti cut a'. flo,, ratc ,! .1 :./.ac. and a subject wasinstructed to keep the crnst-=t ratn )f aif.--w Jf dur-ing the examirr-tion.

Pulmoy,=-y airmvay re:!istance (R' isi g-.ven from the f'ollowing equation.

Where R-~ and R-. =c low re i-a~xe of' the inerted artifIcial resistance and thatof the 7re'oa cFai np~ctively, W~. and 2 are the phase difference bet-ween alveolar pirLu- &nti eifcPt tnme iltuation that the resistance is insert-ed. ori= - can bc ! iV,'I' 1r : 5.fl,/- V. ti'e same airflow (f) rea-

pcc-'.1r, ~ ~,~ 2 .sd:'e.~ i'.'of' b x p;eesure. Pic. 2 show~s a sampletrac-Irg of' he mas!_.,:t

35) The itet)hod ni tie iniinlatioii o)7- d ' ioxC.de.The si)' co .Anirg- ra~, ic:&.c zna-.- by mix'ing With sulfur dioxide gas

to the air aq sno'vw1 in Pig. 1. Te urtntent- of' :,a2Thr dioxide in the air was me&-sared by Rcseilline-Arnaii. ,zethod5 )(.a-tit,-tiv,,,j jvu-t befo- e the inhalation,rd it's q1'-rtT d )L.- this z7hady w~5, 13 .1.0. 20 ppm respectively.

The inhalationi,"t carrd thlc-tgh the moith-p'icce at normal "dreathing rateand mode, when the conte-z-,nata-i air 7sas fliwnn out from cutlo--t on 2. 1/sec.

4J Measuxemont o.' the content of' caifur dioxide ir. the air in the patient room.Corbert of will~r dior':id- in the air in the pstipnt icoor was ireasured chemi-

c~illy bj IRo.an:.ine-or=wdL n method. IDparatlis and priipile was as shown inFig. 2.

The roc. a~ir waa su!okinq' into thE adsorbing bottle with s flow of 1.21/mmi,during 30 :.Lr.uces (36 lit-re in totaCI). Afte2 adsozption of sulfur dioxide in thesolution contp.ining raezrouzic chloride ~2.)and. sodium chloride (11.7%), the con-tent of sulfur lioxide i-a the solution was determined by photomeatry and the origi-nil corcentra' on of sulfur dioxide in the air wao calculated and expressed in pyn.

3r

5) An air purif ier (product of Fuji Electric. Co.) wa. equipped in a patient rcom.The principle of the air purifier in that suspended particulate matters are adsor-bed by the electric discharging method and Liuie y the electric dischargingmethod and sulfurizetl matter3 are adscorbedi by &n acti.vated charcoal, with a filtraration rate of air 10 litre per miiiu-.e. Thi3 Apparatits was functionad only inparticular days.

lie sultsa

1. Variation of airway resistance aui n .-al zubjccts

Results of spirsmetry and Ltiesuroment .f a~irway resistance were listed inTable 1. in the control group, no subjeo! ; had VC of less than 8% of the predicted()value nor P'EVl.O of less than e0% of the r'-dicted value, and Lhe values of airwayresistance (em i{2o,'4/sec) nntnged between 1.5 and 2.4 with a man of 1.85 (S.D.±0.2). This va-iation of' airway r .'1e:.n nornal mibJects seemed to have norelation with sex, age anid vnV1.n'j althoughninru asswssal

2. Airway resistaa.izc in ao.thn-atic aia

All results of ;.ilmnar-. L'Urcti-n '.-ztE h.~ froi asthnatia pa';.ients were list.ed in Table.2, in which severity of' tKCii; z. gc as rild, 1rodsrate andsevere), zkintest by house dust antigen, latest asthma attack anr' other clinicalreferences were also show.n

The values of air.way rosr.ta-i'-o in t-.ism grciip ranged betw;een 0.4 and 3.8 (cmH20//sec) with an aernie C .0 (aD t 3-67). This average value was higherthan the control group:Uase distribution , as pree'mntecl accord, AUnS to airway 1.13istance in different gro-upain Table 3. Values in normr-l co-ItIOl IM3 no~ttly distvrivuted within 2.0 (cm H20/L/sec). Thereforo all cases were divide-I Uinto two groups of' less or over than 2.0.As shown in Table .3, resistanoe cT al~s.'ll of nil. cases were in the normalrange, while mcoe-ite c-tse: -wira mo:tl./ -_' range of over 2.0.

3. Changes of airwqay resistan-o : r -.riaa Lio --f Talfur dioxide in asthmaticpatients and normal syubjects

Hegsurernents of air-day recistzmice ',f ore andi af: er i2.hlaticn of the air con-taining sulfur dioxide were or;.in sixteen asth:-xu_-ic patients and fournormal subjects. All nmbject.3 inhaled the air containing sulfur dic.~dde of 5, 13or 20 ppai during three miaritc.m and apiromsetz7 an i~i eitne anrmntwere followed i-mnmuiatly af'tx', arnd thez. t*hey in-haled again the svrn air Anitherin three minrutes -xd wrr:e testtd again in the same uny. The results were presentecin Table 4, and in Fig. 3.

Airway resistance in the groups (_ E.;thrrtic patients increased partly when 5ppm of sulfur dioxide was inhaled thrf. i v Z;ites, and Lhis tendency was marked whieninhaled six minutes (%Lp. of tw,- succ, .;sive inhalations with 3--5 minutes intervalwith inhalation of hig her lose of t~~:e~and twenty ppm of sulfur dioxide allasthmatic patiento sh±owed isarked incras~e of airway resistance.

On the ofheri-e~nd, normal noj.1. ~. r inc:'-case of airway resistance byinhalation of 13 ppm of sulfur ciiixic'a fcr Lix mirute-, and of 20 ppm for threeminutes.

and norm~al c~abAe ct -

Chnaof the p'nroe-ntagc of FEV 10,, to -the ?Z7 (FFV).t ~ a!~e 4 al.ioio

su1~rur d.:xide in t~~tcpatients -nd in normal subjects were sliowl in Tabie 4and in 71.4. in s;arn casen qtrl.~ rcLip dccrease in Fr ij 0 a~nd in FSV1.o%after inhtlatiLor of' mi1r' ri; oxiiAi* -ii s 3CeA. Tr vi.as.o ip pc'.ld ent corre!atoicnbetween hd ma~iit%:' YI Lh. - oe r-;-d the : d i.P .icxijeb northe der-re ize and th.nn dVo: 4,r n-1 .riat')n. io ,r..t .Au 'he 'ot groupshowed dacrcase in, k.Wl.CAtci~a~cnot~2rdx~c-

5. Daily variation ,f air.,ay r~~sm~ tLat~~tct:CC n'.ii4, trchange-. of' cou',nt of -sulfur diox .dt; in tht!"

Daily charges of airway resistance and FEV10% wre .tollcwcd -nin fuurpatients hospitalized in thn clinic fl.r 9 to 13 dzay3, a dit iii compoared withdaily changes of sulfur dioxdi4e in the air of tha patient room~ besults, wore pre-sented in Table 5. Air.:ay :ienist.:rs:c jr then;e subjcc1.- ...an no,' .tiluenced by thedifference of weather. Thera-fore ele-t~n-h e-~tea~a~rs~tn~and the cont-int of sulu i:cd in t.he aiL. slral 'L'e c!;ipThree of fou~r pattnt s 1-a - ~ st':-' frv;r *h- 'i/.eVal-Je rotlyin rnor-na3. rarge, (uniir c,'! ci H2 )/L/suii ). Cwaau.jt.; a.~ bo.~etween air-way re3i.~tancv- win cz-1tint .)f~~i~ dii.x~:c(e r1jzAt ncL lie evxv;t

Hownv.or, rviticn, Y, :: of slc"-ovn-r~-o - wdv.r ruty of Thc air*c rstti~ vral-ac, Li, Vier-, lrim a ' oii ,: c rZIt~' 'CI'-ei rio ai,;P., r y;r cc1 ~'eco;tcat irn the ai- u rh: vwn in

Thi ainwo .7es i.>riii vi*_n; ir v. '.s :m.:arly un-c:: 2.0 (cm Hi2/Lsee) with~ siall vari~itir reAanr~~: On tor. c'uier-hand, the aiz-vay rerAtsnc:;. c1.g ai,~c ~Lns~sri:t~i:r* wi ranrge.The number Of cases -il',;h airwayi o~i~~'c f m ,re tean 2.0rn 1 iJ 5- ofnild casf'- .;. ,cv;ritr an, 6 ir. 9 of s:?vere cages sggges~.ig crelto

between tz a'~ia r errr stance and th- uv iLv the;d.e'-o±a.-,I ,, -4 F+_n!, ' ' v ad n~icr -f'iisi ± ualfzr d'Lcxide in

shoit na-i .'~ -.,er'~ri~ : i-. *air .±' a.~ :. . 13 anu' 20 p-gza inthe a:ir rii l~ctva.. wie r~.iy) i,',c'~ n_< 3.. 1 -. h 3 pp;-i. Thi zstvggeLs mha n :tr4hxAW*t. lv i . Y.~ ;~.'r t u.7iuUr x.~ by inhalation asto acetylc,-.o14'-w, or l a~re

3.u1far dioxide i the atmosnheric air was t.1-:u4L IIrll ri an. thr'fiipAient even with the c.,mli cf.-,%_. _rc re J~ is~ j

M.e airiay re '4stance varied evter~r lay in licu:' datient a during '- period ofthi s study. Three if them were mostly rernal"iing in norial range Only onepatient showed marked variation dlir71.ng the p iriod re.flectinri the con-tent of rulfurdioxide in the air. This patien~t shc~t a positivi-: cer:'&lation Ibetween his airwai-rresistance and atmospherie, wu.fur diox'-.de content Thiz- is a goou' ."vidence thatsulfur dic-dide in the atrnosphvric air ...nfluences to the obstractivr. di-turbance inan asithmaitic patient.

4. In the inhalatic-, test of mtxlf'.r diodid in short perl-d airway resistan-.e ofas&.hmatir; patients increaosed *.i!i.y,-,hi e KW).. % ot came pa .inrts ! Iowed no

einiiaz' cn'ge Tisi.nr-ably duv to the fact th!tt t!!-:, in.T of airi.way resls -.r.:e is n.-.e -erc-~tivn~ t-n L'.he ofxr.~nLcI FFV*. -hihoo-kij.- t .1,cops:,atioL of the :iubiiit

Sumary

1. Fifteen normal subjects and eighteen asthmatic patients were tusted by spiro-graPhY as well as body plethysmography, and ehanges of airway resistance beforeand after inhalation of sulfur dioxide in three and six minutes were followed andanalysed.

Variation of airway resistance among normal subjects ranged between 1.5 and2.4 (cm H 0/L/sec) with a mean of 1.85 - 0.2 (S.D.), while in asthmatic patientsit ranged between 0.9 and 3.8 with an average of 2.03 ± 1.67 (S.D.). Airway re-sittance was higher in moderate or severe cases of disease than in mild cases.

Asthmatic patients showed increased sensitivity to sulfur dioxide at 5, 13and 20 ppm in the air during the period of three and six minutes inhalation -whileno change was recognized at 13 pin in the group of normal subjects. FEV 1.% asth-tic patients showed no change when airway resistance varied markedly.2. A correlation between daily variation of airway resistance and daily changesof the atmospheric sontent of sulfur dioxide was noted in a mild asthmatic subject.

0

References

1) Oshima, T., Ishizaki, T., Miyamoto, T., Shimizu, T., Shida, T. and Kibe, T.:Air pollution and Respiratory Diseases in the Tokyu-Yokohama Area, The Amer.Rcv. Resp. Dis., 90, 4, 572-581, 1964.

C 2) Oshima, Y., Ishizaki, T., Kiya-moto, T., Kabe, T. and Makino, S.: A Study ofTokyo-Yokohama Asthma Among Japanese, The Amer. Rev. Resp. Dis., 90, 4, 632-634v, 1964.

3) Mochizuki, M.: On the time relation of airflow and alveolar pcessure, Kokyuto Junkan (Respiration & Circulation), 9, 149, 1961.

4) Mochizuki, M.: A method for measuring airway resistance in man, Kokyu tojunkan, 9, 157, 1961.

5) Comroe, J. H. JR., Botelho, S. Y. and DuBois, A. B: Drsign of a Body Plethysmo-graph for Studying Cardiopulmonary PhysioloU', J. App]. Physiol, 14, 439-444,1959.

6) Kanagawr Pref. fleport: Rosanilin-F raslin Method, Annual Report cf Air Pollu-tion Sui vey (1957-1962), 3-4-1, 19(2.

7) Makino, S.: Bronchial Senitivity to Ac!tylcholine and Histamine, J. Allergy,38, 3, 127, 1966.

-6-

Pic. I Apparatus for Ibasmr t of Airifay Realatanc*

A subject sits inside a body-pletb7mograp*h with its plasticdoms open. The body-plethy sgaph oan be mde airtijbt by! oloelnte dc.ms.

all A I

~BOX-PkES SURE _t'

Pic. 2 Simultaneous Recording of ktrf~x-i ard lo-r-" esari

During recording on the 2..f sirte oVr thie arrcw an aztici"1r t; lwas removed, and during recording on the right si~c i':.e r~ .- wsertod. Airway resistance can be ca'.cu~hted frc, tini :'-atio of V- to '7,Incase f1 is equal to f and the values of from~ ro~IsLI=c oL M ati-fi-calresistance and pneaoachograph ane known.

Air.~mLu fig T.Iti of SA~W fm

OA 6

Fig I The vL4J1 mockafdm of sulfutimdA4 lUte air wAn ialationtchiu'q"a of mix the air to mn

mI"w#AwAO

9 -77,

Now-..

Fig. I! The mesu~remnent of the concentration of sulfur dioxide it the airthe, atmosheric air Of a patient-room.

10

It

I ,

hii

-- 4--<

0 3 6 - 3 6 3 6

Fig. III Airway Resistance on Asthmatic Patients responded to the InMIaion

of Sulfure Dioxide Gas

- Patient, N-- ormal Subject

. .....

D' onq of-MGrm0 S*f kj

'Pat5ientS, 3...a Sub-lee.. ,

1012

Table 1, esul.ts of Uan Function Tests on Nozml Subjects

Sax Age VC. FVC 7 1.0 % FEV 1.0 AI~imy(cc) (cc) (cc) lResistance

S.I. M 16 4150 4000 3680 97.5 _1.8

Y.A. M 16 4400 4320 3880 89.5 L2.4

Y. S. m 17 3810 3680 3440 94.0 1,3

M.N. 417 150 3960 3650 192.0 1. 6

M.S. M 19 3950 4180 3820 91.0 1,9

M.H. M 26 2.0

M.T. M 29 4120 3860 3400 90 1.8

H.N. M 29 4000 3950 3360 85 1.5

1 S.T. H 29 4380 4.280 3540 82.5 1.8

s.N. 3 50 7.5 I - 1__________

T I.M 0370 360 3040 84,.0 !, .8

F .S. _ 22 3410 3440 3060 89.2 i_1.9

II

R.Y. F 42 1.8_, _ .I _ _ I _ _ I _ _ _ I _ _ _

A,70,ae /PSx C.Z

13

I I

-a 0 -A

*4,)

0 00100 0 x t o~ 2 O a. A: 00a2la

ig o a a " 44 to o 8

t-- 100 (V 4 ; p

0,r

F4-

C -C

10

~~~~C~. I-o Xid

+1+ + + + +

0 0 0 0*

x x4

.4 PI.. . 4 .

. ..

Table 3, Case Distridutio, of Airway Resistance between Different Groupsof Severity

0

a everity Vf Disease

Ml Cases Moderate Cases

Less Than

2,0

Over Than

~2.9

NoteI One of Severe Case w inc±iuded n tho Colin cf Noderate Cases.

15

TAble4. Changs of Airway Resitance !.±ter Inh tlion of Sulfu Dioxide inAsthtic Patients and Normal .ib~ects

Sbe xo Contert -rwes1.stan,," FEv 1.oof Before Inahaiir erd Weore lraltin Priod~

Asthmatic i 3 3 7z oi~n, in, 6min______ _____ . o a i ____ .__

G.A. N ,2 13 2.8 . 5 -76.0 ,1____ 69.7

T.&! F 6 2 2,4 Z..9,. !,. 63. 5

N.O. N 2 5 .0 . 90.0

ss. N1 2, 5 1 .o 2.5 7. ...7e.O . 71. 0 69.1

SIS. x 35 5 0.9 t 10 2..). 81, 2 3.t 89.2

S.N. N J0 5 0.8 __..__ _____;_

Y. . N 50 1 " 1.3 1.7 .63,0 58 61.7,,,______ __2.9____ 63~ 0 6. 6h

55 3. 1 .4

_ _ . 8 2 20 1.5 1.7 12-,- . 83.0. 29 20 3 , ; , 2 . -1 - 67.5

AK. N 29 20 1.8 5,4 I 6

K.I. M 32 20 2.2 3.7 .6 5 5/15 54.,8 4

H. 0. 5-35.20 3.0 5 L, 2.,2 54.4S.". N I -2 7 68,2 i

M.N. F 2 L5 23 2,. i 47. , 56.5....

Nor-=".

N. 29 33 1,6 .. ! : 7,' 8.C .

V.?. K 28 13 1.3 I1.5 1. 91.0 1 88.5

I ~.-1_- .J 3 ~ . S .C80.1

T.3 , F 22 2k 2.0 2, 3,,-, !j9.',. 91_5 1 -. 5

16

. . .... . .. - . . . .. II I ll

PART II.CHANGES IN MECHANICAL .0vTERTS OF GUINEA PIG IN EXPERIMENTAL ASThXA INDUCEDBr ACETYLCHOLTNE OR HISTAknuz

Since the shock organ of guinea pigs is lung and inhalation of aerosols ofhistamine or acetylcholine causes respiratory changeb .esembling to the humanasthmatic attack. Guinea pigs has been uses to investigate the offects of variousdrugs for bronchial asthma. Changes of ;aaohar.lal vroperties of the lung in ex-perimenta4 asthma was reported almost identical to tnubC .n huran asthiiaticattaks.2 ) However, the changes in relation to doses of a -not been well studied.

The aims of this study are (1) to know how mechanical propertien of the lungof guinea pigs change in relation to doses of administered histamine or acetyl-choline, (2) to know which of measurement will be appropriate to represent experi-mental asthma.

METBODS

Guinea pigs was anesthetized with ether and a polyethylen catheter 1.2mm D.M.was introduced into the right middle intrapleural space, and placed in the body-plethysmograph according to Amdurs method 1 ) as shown in Fig. i. IntrapleurRlpressure was transmitted to a ressa transducer (Sanborn Koicl '26S) throughthe intrapleural catheter which was filled with saline solution. Ai-flrw wAZ.e&.3uroU ..-mh a meh flowmetoz att.achad xth plastic facemask and Statham differ-ential strain gaige (Model PM 97). Volume was calculated by electcl integra-tion of the flow signal %ith integrator (Sanborn Model 350-370OA). Pressure inthe bodyplethysmograph, i.e., pressure applied around the chest of ' guinea pig,was measured with Statham differential strain gaige (Model PM t0.5). Bodyple-thysmograph was connected to a oscillating pump which can supply s tnewavc pressurechange of 18 C/S. These signals were amplified with Sanborn Carrior Amplifiers(Model 350-1OOB) and recorded on direct recording sistems (Sanborn Model 9641) ordisplayed on C.R.O. (Sanborn 569B Visoscope). Pulmonary complione.ce an-! y;!_,onaryresistance were calculated on recorded tracings following Neergard and irzinmethod. Total flow resistance was read by loops of flow and bodyplethytmographpressure displayed on C.R.O. which were photographed ,-- Sanborn care.tr' followingMead's oscillating method modified by Swann, st. al., as described this semi-arual report.

Minuze volume, work of respiration, frequency of respiration and ratio ofexpiriunm to inspirium were calculated on the recorded tracings. In some expari-ments we mearared approximate changes in chest volume using a pneumogre.ph a-roundthe lower thorax.

After recovered from el.ther ansthesia, aerosols of histamine or acetylcholinesolutions was inhaled to a guinea pig for 3 minutes. The concentration of inhaledsolutions were doubled serially, ranging from 0.16 to 20.OmK/ml of histamine andfrom 2.5 to 80.0 mg/al of acetylcholine. Measurements were car.ried out immediate-ly after the inhalation. In case pulmonary compliance decreased after a inhala-tiov, the next inhalation was postponed untill compliance value returned to thestandard value.

17

, - I

'k , :. . , , .

.- SU9LTS AND DISCUSSION

fig. II shOWS Changeae in( aMoneary oMpliance,(CL)pulmonary resistance,(RP)total roep~ratory resistance, A)irk of breathing, respiratory frequency, minutevolume. ratio of expirium to xpirium for various concentration rf inhaled hista-mine. . RL and 'I TIncreaed abruptly with histamine inhalation at the time of shock as much as twicethe control valus, while they presented no definite tendency ihange, fluctuatingwound the control values, until shock occured. Though animals presented dyspneicappearance by increasing dose of inhaled histamine, there was dound frequently no&efinite increase of resistance, both pulmonary and total. Guinea pigs died some-times in shock when the resistances increased as to double of control value.Pulmonary compliance began to decrease on inhalation of histamine with the concan-tration around one sixteenth of shock concentration. Beyound this concentrationalmost linear relationship was found between CL and logarythm of histamine concen-tratio r. Since compliance beean to decrease earlier than other measures, it isconsidered to be giod measurement to represent pulmonary sensitivity to inhaledhistamine.

Work of reapiration increased gradually in proportion to decrease of CL. and0Increase of R.L Though minute volume was kept constant in spite of increasing con-e49tration of inhaled histamine, it began to decrease at the inhalation of shock

ol of that. It is considered that decrease of minute volume or increase of workof respiration is appropriate to represent dyspnea of experimental asthma, thoughfurther comparative work will be needed to clarify the relation between dyspneaand changes of pulmonary functions during the asthmatic attacks of human.

Frequency of respiration changed in preportion to the changes of time con-stant, e.i., product of CL and RL- It was maximam at the concentration of I orof shook dose, and began to decrease in accordance to the abrupt increase of R.

Ratio Exp/Inep increased remarkably at shock, which has been used as a index ofexperimental asthma.

Though among the measures observed, pulmonary compliance showed the most con-sistent change for increasing dose of inhaled histamine, its measurement needs tomeasure the intrapleural pressure the procedure of which hurt animals and conse-quently on lon-tars experiments CL with this method can not adopted for succesivemeasureme:t with relative long intervals. In this respect total flow resistancemeasu-emcnt is considered to most appropriate to express experimental asthma,since it increases at shock as definitely as easy to detect and is suitable to beused on long-term follow-up study because it dose not hurt animals. While RLpresent the same change as RL, it can not be used because of the same reason in CL.Other measures such as respiraton frequency and Exp/Insp ratio changed at the nearshock, but their ohanges did not appear difinite enough to be the index of experi-mental asthma comparing to those of RT.

Fig. III shows the change of mechanical yroperties by acetylcholine inhala-tion. The methods and presentations are identical to those on histamine inhala-tion. The results obtained on acetylcholine inhalation were almost similar tothose on histamine, though an impression was given that resistance began to in-crease earlier than in histamine inhalations. Resistance began to increase onacetylcholine inhalation with half of concentration at shook. This result mightsuggest that in experimental asthma of guinea pigs acetylcholine cause constractionof the bronchi more easily than histamine do.

The concentration at shock ranged fromflomg toftmg with the mean of f33 inAch and ranged from2O mg to 5 ug with the mean of /4. in histamine. The ratio ofmear ach concentration to the mean histam ne concentration was aboutlOsL. Thisratio was identical to that .observ in asthat patients which bronchial sensi-tivity was measured with the mniml 6ois to oem 10 percent decrease of FEV 1.0.This finding was interesting to suggest that pharmacological effects of histamineand ach might be similar in the lung of human and guinea pig.

16I

Fig. IV shows changes in chest volume, compliance and resistance in relationto the concentration of inhaled ash. Sometimes resistance decreased temporarilyon inhalation at i or i of the concentration at shock during the course of in-creasing concentration. In order to understand this phenomenon, as speculationwas proposed that this decrease in resistance might come from oompensairy in-crease in lung volume since resistance decreases in large lung volume.+/ Contraryto the speculation, no increase in chest volume was observed on the inhalation inwhich resistance decreased. In this connection Nadel stated in experiment withcats with histamine L.jeotion that the increase in pleural pressure and decreasein complianc will tend to dilate airway and this may account for the decrer.se inresistance.2 / Further study will be needed to clarify these problems.

Vig. V showed correlation between pulmonary oonductance(/R) and total con-ductor (1/PT. Correlation coefficients were 0.64 in hist. and 0.69 in ach. bothwere statistically I significant at the level of 1%. This results suggest thatmeasurement of RT could be used for that of RL-

SUWARY

Change of mechanical properties of the lung of guinea pigo was studied in re-lation to dose of administered histamine and acetylcholine. Total flow resistanceand pulmonary resistance increased abruptly at the time of shock as much as twiceof the control value, while pulmonary compliance began to decrease on inhalationwith the concentration around one sixteenth of shock concentration and thdre.teralmost linear relationship was found between pulmonary compliance and logarythmof histamine or acetyloholine concentration. Since the measurement of pulmonaryresistance and compliance required intrapleural pressure and this procedure hurtsanimals, measurement of total respiratory resistance is considered to be appropri-ate co express experimental asthma in long-term studies.

£9

wmom""

1)And=*z X. 0. *t .1.: Mechanics of respiration in uwnethetisd guinea. picotAmer. Jo Pb~miol.; 364p 1958

2) Colebutch, E. J. No ot al.s Effect of histamine, seratoaiin and acetyleholinean the peripheral airways, Jo Appi. Pbysiol. 21; 217, 1966

3) Stein, N. &t. &1.t The Mechanical properties of the lun~g in experlomtalasthm in the guinea, pig J. Aller~ro 32; Op 1961

4) Swum, fi. E. et. al., Pulmonary resistance measurement of Suinea. picot ANAArch. IEiviron. Health, 1C; 24, 1965

20

Fig. I Photogr aph of Apparatus.A guinea pig was placed in the bodyplethysmo&xTaph. A polyethylencatheter was inserted in the Pleural space and connected to a yrtsauetransducer. Airflow was measured b~y a plastic weah1-flowmeter. BoxPressure was picked up through front t'.ibings Oollatlfl3 wessttre wassupplied from a Pump through a rear rubber tube*

21

-A K

% of %t.t I

0 "", \ "' d Z/Mo

O F h i

\ Umc%__

I , *N //

, of , o.trO: i/ . ,

; , \ / / 2CC-"

o - i /" . / -° / '

I '7 CCKKz2.

E, ,:,C .'IT -T, .A CFhii$ .-.v:',

Fig. IIChange of Pulmonary comliance, Pulmonary Resistance, Total Respiratory Re-

sistance, W-nute Volume, Work of breathing, Frequency of Respiration. Ratio ofExpirium to Inspirium in Relaticn to Various concentration of Histamine Solution

Measures wre expressed as percent of the control values. I'he concentrationof Histamine was expressed as the ratio of concentration where a guinea pig pre-sented shock.

22

77 1 _ _ _ _ _ _ _ _ _ _ __ _ _ _ _ _ _ _ _ _ _ _ _ _

too,

WORK Im

WS?'-:EITOT

ftCSISTXUCE

/2 . /2

7. of ew.'fCQNCEN?-AT!ON Ofr ACM

so~

COV*4AT1')4 Of AC"

Fig. 7m MlangP Of ?"-)1n~narY com~lirtnce, Pulmonu7r Resistance, Trotal RespiratoryRe313t.W 9.K-~ Volume, Work of breathing, Freouency of Respiration.Patio C' Ezi -- , to Inspirium in Relation to Various (jonceztration of ace-

w ~c-e ex-ressec 9. percent of the ;ontrzol values. The concen-trstion -.- Acetyrlcholin~e ru expreoed as the 1z1io of concentraticz wherea guin"&. pig prepented shock.

C) 3

AHCHEST VOLUME 0 0 % fCnr

40P,/Z =

20 R

10 RL

50 tFZ2.ZZf

Fi.IV Change of Chest Volume, Pulmonara7 Resistance and Pulmonary Compli-ance in Relation to Concentration of Acetylcholine Solution,

Approximte Change in chest volize was measured by pioumographattached around the lower chest. in inhalation of 4ong/ml. of acetyl-choline where resistance decreased by 30% of control value, noIncrease of chest volume was observed.

24

Iree

- - 3

-CASES 14 -CASES -

to 00 .4 , Cfofite to MC~t

I / Total Resistance I/ Total Resistance

HISTAMINE ACH

?i4. v IsUtiou-ahip botwwen 1/ita~y D.esitoan 1 3/Tota Romp1:tory

2-5

-nor OWsuLymm DCIamosoU To LCUT L ASTHMU i UInEA PIGS

ifUre dioxide In the aftopbore of urban area, has been considered to berelated with increasing frequency of asthmatic attacks. In order to investigatethis zalatiua, influence of sulfure dioxide exposure hUe been studied in variouseaamlse In our previous report sensitivity of the bronchi to settyle line wasInvestigated in guines. pips exposed to sulfur. dioxide and not exposed. Thoughthere was found no significant difference of bronchial sensitivity to acetyihbolinebetween tbem, conclusion has beon veserved since experimental asthmtic attackwas observed only by the change of alveolar pressure pattmrniand this method wasnot quantitative enub.

In-this study experimental asthma was quantitatively chocked by the increaseof total flow resistance. This paper was aimed at knowing whet~her, in guinea pigs,

0 the exposure to sulfure dioxide accelerate. the experimental athma, induced by the

MATERIALS AND XTSODS

MATEXALMae guinea pigs, weighing 250 to 300 go were used.

-UTEMSi) Measurient of total respiratory resistance

Total veopiat=7 5esiatance was measured by medts oscillation method modi--, ied bY Swami St al.. The details in measurement-was reported in SeimiualReport of l.965-35 The principle is as followas An animal wes placed in body-PletbYsmogrpmh in which sinewave prea~surs of 18 cycle per second wasn appliedaromd its chest. Total respiratory resistance was calculated from the ratio ofpressure Change to resultant flow change,

C) ~ii) Mesurement of %gpchial sensitivity to itieAn desoribed In 8an alreport of 19650 oha sensitivity to hista-

nine wsepsdas AN , i -e., the dose oftbstamdue to caus the increase oftota respiratw iestance donblo of the control Value. An animal mas allowedto Inhale seroup1 s Of hitftin* in9asing their 000cintraticu until apparent shookwas observed. 14 =6 Lculatod from *M dose-reepose curve.

iii) ktpours to suitos dideGuieaO "& were expo~ed to muUfMr &ijdA V4~ OW tratione of 20 and 2W0

ppe for the perio ot am hour. wAeuroimt ofssitivity to histamineW" carried "ut 1) imediately after the first=%V:r=* ups0 doy after the lestexpetue a Is" 4al3, meectivo expo"Me. r~barpoar biterr U 4 was suppied fron1.0 percent idadde es.~o M its eaMoatatlM "S asasurd IW beaSLimin

RESULTS AND DISCUSSION

Piq I and III show bronchial sensitivity to histamine before and after sulfurdioxide exposure. The mean Hi of f ,.' guinea pigs was: 3ZxrgJ/m before the expo sureand after the first one hour exrosure the mean H was 4mxJm! and after the sixdaily consecutive exposures the mean 91 was 10mrg/ml, whereas the mean H of f'.-.controls were3$)4,61 , and 90 m/ml at each coresponding occasions. The change ofHlafter sulfur dioxide exrosure was not statistically significant comparing tot at of control group. The mean Ei ofe. . guinea pigs was 3.15mg/rI before the ex-posure and after the first one hour exposure the mean H was 3.46n /m.lar.d after thesix daily consecutive exposures the mean 11 was 7.(,2.mg/?@ whereas the mean HE of

oi . controls were 3.1, 3.22 ant. 66mg/el at each, corresponding occasions. Thechange of H after the exposure was not statistically significaut comparing to thatof control group.

These results showed that exposure of sulfur dioxide did not influence bron-chial sensitivity to histamine in guinea pigs, taking th apparent increase oftotal respiratory resistance as a index o experimental asthma. They ooincided theresu]ts reported in our previous report. 2 ) As described in our previous report,the polluted atmosphere of the Tokyo-Yokohama area increases the incidence of re-spirsto.y syri:)tors such as coughing, apatum production, tlhzoat irritation andpulmonary function disturbances, while there was no increased incidence of airwayobstructive disoases.l)4) It is considered from the results obtained in this studythat Larpollution including sulfur dioxide mlght have little part in the ethiolo~rof bronchial asthma, though it causes symptoms in asthmatic patients as shown inthe following section of this report.

%bo inflnce of sulfur dioxide exposure to xperimentvl asthma was euiedin guinoa pis, measuring bronchial sensitivity to histwrine by the .a.-wsg of totalvespiratory resistance. No vignific~nt changs of bronchial sensitiv-1ty to hista-mine was observed amorq the groups exposed to sulfur dioxie, comparing to thatnot exposed. The results gzive us cn impression thAt tle air-yo1Dution v1: 1ht be oneof the agravating factors of bronchiai asthma but have little role in the etiolo-

27

_.. ... ... ... .. I I . .. .. . . .... .. .

REFERENCES

1) Imhiaki, Tooa Studies of respiratory diseases due to air pollution in Tokyo-Tokobam area, Final Report, Contract No. DA-92-557-FEC-36323, 1963

2) Ishisakit T.,s Studies of respiratory diseaes due to air pollution in Tokyo-Yokobsm area, Final Report, Contract No. 12-557-EC-373O8, 1964

C)3) Ishisaki, Tops Studies of respiratory diseases due to air pollution in Tokyo-Yokohama area, Semiannual Report, Contract No. 92-557-PEC-38659, 1965

4) Oshima, Y. et al-I Air pollution and respiratory diseases in the Tokyo-Yokohamaarea, Amer. Rev. Reap. Dist 90; 632, 1964

~)Swann H. 2., et a.9. s Pulmonary resistance measuarements of guinea pigs,AMA Arch. Environ. Health. 10; 24, 1965

28

w Shock

I-~ % ofControl

C,)6j

wM.2

CI.-

.L- * 63 12L5 2-3 50 100t420.0Le0

CONCENTRATION of HISTAMINE

Fig. I. change of Total Reupiratoy Resistance in Relationto conentrtioni of Htistainen Solution InhAled.

Bzonchial sensitivity to histamine was represented bY H&, theconcentr'ationl of histamine solution whih was asmmd to be requredto --s 200 percent increase of tota respiratOrY resistance Onthe dos response rslatim wne.

29

se a 1 04 m

-1. 12

d) 100

A £cotro / B cotro

@41 033

Ou 043

10350

g .1 200(g..., 0.CA MWf EV E.#- N MI. f.,

Fi.I. Cge of Brnha',.'iiy o -. ai fe

(right Exom f3Opm

A ddSWOosd 13 Eposed

0443

E

Aft Go BOWSo C Aft GoSSn

043 O.w

2w ,. s ,m[.

8.."U Af, t

Fig. III. Change of Bronchial Sensitivity to Histaminmafter 7 Daily Exposures of Sulfur Dioxide

(left) Exposure of 20 ppm,(right) &qposure of 200 ppm.

31

Awaording to thoe results, the influence of sulfur diox~d exposureto e remzental sath-a was studied in guinsa pigs, nasuring bronchial snsitivitytO histamine by the change of total respiratory resistance. However, n significantshanme o bronchial sensitivity to histamine was observed amng tUme aoipa exposed.Th. results give us an ipression that the air-pollution" might be one of the aggra-vatig factors of bronchial astlaa but have little role in the etiology. (Author)

4

p,:

• - P

Unclassified

"D = FCOMU IT O ATA. - M

Departmnt ofIyit hrp eiieUcasfeUniversity of Tokyoj, School of Medicine, pTokyo, Japan

S. "IPONT TITLE

R3SPIRATORY DISEASIS WUE TO AIR POLLUTION IN TOKYO-YOKOHAMA ARIA (U)

Final iteport No. 6,30 September 1965 - 29 September 1966IAurh@~) 66"o* own am#RM0

*Ishizaki, Tatsushi

4 0161000 "ATo is. VeYI.uSo P *g. , we. op mapa d

Ja~mary l967 1 =16

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06 A VA %.AWPTJYY/UTAYUP U?8C3

Distr ibution of this report is unlimited

It0. W 6 AOTA *41WC S& N0S UTAav A91W1?IVU. . ApqR&D Group (Far 7"t)

III. AGSTIACV

a~s*.J- Fitee noml ubjctsand eighten asthmatic patients were testedby sprgah n oyplet1lrsmographys and changes of airway resistance before

adafter i nhal1ation of sulfur dioxide in three and six rinutes were followd and

-- AI ma offraino airway resistance w~as 18 nnrlajects and'23 in asthmatic patients, and S.D. of variation of that was &0,2 in

forar nd 637in latter. Airway resistance -was higher in mdrt rsvrcases of disease than in mild oases.

Asthmatic patients showa increased sensitivity to sulfur dioxide at'5, 13, 20 ppm in the air during the period of three and six minutes inhalationwhile no change wg -ccn.ized -+ 13 ppu" in the group of normal subjects. Acorrelation between daily variation of airway resistance and daily changes of theatinsplwric content of sulfur dioxid was noted in Mtastheatic subject.

P41P. In experimental asthma, change of mehanical properties of the lungof guinea pigs was studied in relation to dose of administered histamne and

aceyloolie.Total flow resistance and pulmnary resistance increased abruptlyot the tim of shock as much as twice of the control value, while pulmonary corn-liance began to decrease on TiAltion'with the *oncentration around one sixteenthof shock concentration and therafter alIiost lnar relationship waa found betweenpulmonary compliance and logarithm of histamine or acetylcholine concentration. )

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