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Methods of Procedure I n Stage I1 we plan t o use the same site and species b f animal as f a Stilgee.f ; each animal w i l l undergo the same pre-exposure tmtraent- anesthesia, paralysis, and mechanical ventfaation. will be attached to the shaved skin o f the animal, The point sources w i l l be positioned i n a triangular conflguaation so as not t o block the'Anger camera's vlew o f the respiratory systerrr and still be nithln the camera's overall f i e l d of view. The animal will then be supported I n an upright positlon and anterior-posterior and lateral grays will be taken. The X-rays will later be used to superimpos&the anatomic boundaries o f the respfratory system onto the scintigraphic pr ln dt, usfng the basic techniques o f Coulam --e tal, (1) The anbal will be supported fn an upright position throughout the experiwnts, since gravity appears t o be the principal bassis of uneven distribution of ventilation and perfusfon. The following measurements will be made I n Stage 11: Ozone uptake using 015 labeled O3 FOLDER r!I A Lung Function measurements will a1 so be done, using the best method(s) determined i n Stage I for &e detectfon of early O3 caused alteration in lung function, The lung function data obtained will l a t e r be compared with the Anger camera data t o determine the sequence and possible interrelations of CY- The&tagged gases C02'5, N 2 1 3 s and Co* will be adtninjstered using t h e saw stetptyped method. The animal w i l l be hyperinflated by clamping the 6 ' 'I_, . t . :a52
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exp fred air pathway for four sequential breaths. tracheal cannula will then be & switched frorrr the O3 pm supplying the tagged gas. The tagged gas pump will be The Inlet tothe hed with the tagged gas and operated a t the same frequency and tidal volme as the O3 seconds and the npump, The animal will breathe the tagged gas for - be transferred back tothe O3 pmp. The camera system wfll record count rates -R?BEH frora the lung field during tagged gas uptake and washout. Data will be 7BZS€ll stored i n fields containing one Sns pi ration - expiration cycle, so that later breath-by-breath analysis of the dab may be possible. Multiple breath of the tagged gas are necessary t o achieve acceptable 1eveb.of positron activity for the Anger camra when it f s operated ina cofncfdence detection mode, Paul Meyer wfll add the following sections; Camera description - operation and problems si Data analysis Erv's article on data acquisition from the camera g' I will add sections on de tenn f ning + V / Q ratios and diffusion rates, Sftes of 0 uptake cfll be determined by exposing an animal who is breathing air (and GO2 if exercising)to 03. using the same equfpment described a{iG'prt. The exposure nust be* of cancentration and length necessary to obtain suf fi cfb nt positron events with the Anger camera for later data analysis (minimum -e vents). We hope to eventually use a n O3 concentration of @J 0,l to 1.0 pp. The relative sftes of O3 uptake will be determined by the number of positron events at any c ne site within the res pir sts ry systm),the sites with the highest activity befng the sites of
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ufll then be superfmposed on the cwra printout so that ident f ffcc t fm of approximater 1 sltes o f uptake can be made. The experiwnts will he done on aabiar’s both at‘‘ rest and during exercise (double-nestfng mfnute verrtilatfon). the s m technique as f n Stage I , They wlll be repeated once every 20 minutes throughout the contmf perfod and exposure. Ozone exposure and C02 levels wtll be controlled usfng exactly the sac methods described fn Stage 1. total O3 exposure wSll be three hours witha one-half hour control perlod (air) preceding the exposure. Ue wfll again start w f t h a concentration of 0.4 pjm O3 and ’lower the concentration i n succeedfng groups of animals until RO sfgnf ficant a1 teratfons I n regional i/Qand/or Co* diffusion are evident. Preliminary tests a t a test all procedures and treasureweb. The measurwnts will be clone on four groups of animals (approximately eight animals per group): 1 ) those withm exposure t o ozone (rest); 2) those with no exposure t o ozone (exercise); 3) those w l t h exposure to OLOW and 4) those with exposure to ozone and C O P (to siwrla te exerci se by hyperpnea).
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fiethods of Procedure In Stage I 1 we plan i o use the sane size and species of animal as i n Stage I ; each animal w i 11 undergo the same pre-exposure treatment- anesthesia, paralysis , will be attached to the shaved skin o f the anfmal positioned f n a triangular configuaation so as not t o block the\ Anger camera's The point sources will be view of the respiratory system and still be nithfn the caraera's overall f i e l d of view. The animal will then be supported fn an uprlght position and anterfor-posterior and lateral grays will be taken. The X-rays will later be used t o superimpos&the anatomic boundaries of the respiratory system onto the scintigraphic print&, using the basic techniques of Coolam --e tal.(7) The anha1 will be supported fn an upright position throughout the experiwnts, since gravity appears t o be the princlpal bassis o f uneven distribution o f ventilation and perfusion. The following sueasurements will be made i n Stage I?: Pulmonary perf usian using C02 Pulmonary ventflation using Np Lung Function measurements will also he done, using the best method(s) determined i n Stage Ifor fie detection o f early O3 caused alterdtion In lung I function. The lung function data obtained will later b e coinpnred with the CI Anger camera data t o determine the sequence and possible Snterrelations of saw sterptyped method. The aniriial will be hyperinflated by clamping the
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expired air pathway for four sequential breaths. Tke inlet t o the tracheal cannula will t h e n be & switched fro3 the O3 purrig t o a pump supplylng the tagged gas. The tagged gas punp will be pre-fTushed w i t h the tagged gas and operated a t the same frequency and tidal volume as the O3 seconds and thenpmp. The animal will breathe t h e tagged gas f o r - be transferred back t o the O3 pmp, The camera system will record count rates %!S6# from the lung field during tagged Gas u p t a k e and washout. Data will be stored i n fields containing one Snspiration - expiration cycle, so that later breath-by-breath analysis o f the data may be possf ble, Multiple breaths of the tagged gas are necessary to achieve acceptable levebs of post tron activity for the Anger camera when itis operated ina coincidence detection mode. Paul Meyer will add the following sections; -,. Camera description - operation and problems Erv's article on data acqufsition from the camera &' Sites of 0 uptake cf11 be determined by exposing an animal who i s breathing a i r (and C02 if exercising) t o 03. using the same equipment described i n the C0215, N 2 1 3 , a{CO*'worko The exposure nust be* o f concentrzition and length necessary t o obtain sufficient positron events with the Anger camera events). ife hope to eventually use an O3for l a t e r data analysis (mininun -cr determined by the number o f positron events a t any cne sfte within the respiratwy systemtttie sites with the highest activity befng the sites of Oolto 1,O ppm. The reitative sites o f O3 uptake will be
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highest O3 uptake. The X-rays obtained i n fie beginriing will then be o f uptake can be made. The experinents will he done OH amfnals both a t rest and during exercise (double-resting nfnute ventilation). Respi ratcry function measurenants will be done m plo ying exactly the sa,w techniqu& as i n Stage 1. They will be repeated once every X!einutes throughout the contrcl wried and exposure. Ozone exposure and C02 levels will be controlled using exactly the same methods described in Stage I. Total O3 exposwe will be three hours wit4 a one-half hour control perf od (afr) preceding the exposure. ble will again start w f th a concentration o f 0.4 pprn O3 and lower the caocentration i n succeeding groups o f animals unti 1 no sjgniffcant a1 terations i n regional V/Q and/or Co* diffusion are evident. Preliminary tests at a concentration o f approxltnrately 1 p p i will be done on tno t o four animals to test a l l procedures and Rieasurem&. The measurements will be done on four groups of animals (approximately eight animals per group): 1 ) those w i t h no exposure t o ozone (rest); 2) those with no exposure t o ozone (exercise); 3) those with exposure to ozone and 4) those with exposure l o ozone and C02 (to simulate exercise by hyperpnea) .