Tobacco Documents Online

Ttlli C(1L'N(9f. 1''UR TOI3ACCO RESEARCH-U.S.A., INC. I Septernber 8, 1981 i MEMORANIDiIM 10: Dr. S. C. Sarmexs and Staff F1mM: D. H. Fbxrl and R. C. Hockett II '1 ~±+.: ! (1 .1 I).~ SUBJfX.T: Site Visit with Dr. J. E. Repine and oollaborators at the Webb Waring Institute, University of Colorado, Denver,l]olorado, August 24, 1981 Grant No. 1322M "Basic Mw-t,anisms of Lung Injury fran Inhaled Oxidants: Elucidation of the Role of Hydroxyl Rsdical. (Qi) using Dimethyl-Sulfoxirle MtSO) " Dr. Iiepine introduoad the work which has been ,+nclP*+akp*+ by his gcoW over the past eight months sinae the start of the grant, which has to a oonsiderable degree been eowered by their most recent progress xeport. Subsequent piesentaticros then eovered individual eouponents of the pzogram by the individuals ecmcerned. The over- all oancept is that free . Ckl radicals generated by activated PNIlVs are highly to:d.c and contribute severely to pi7mo+A*-y disease. They have then utilized various sca- vengers to prevent the lurkg disorders. R. FOX: (Support fran NIH for five years) 7he approach used by Ftiox has been to enPloy an endotoxin lipopolysaecharide (I8S) which appxears to protect the lung against exposure to high levels of mygm, alttmugh it had ve•sy little effect an the n:mber of P.4PLs reoovered in lavage fluid. Howiever, it did seen to protect against the increase in cell population turnover of PANIs and the pailsnonary injury caused by high oxygen levels. F1:rther, alpha-naphthyl esterase actiwity, which is elevated by high oxygen levels was lower in the IPS-treated group. Ox}+gen damaged PAMs release che:notaxins for PmAis. LPS appears to prevent release of these chenotaxins'artd to therefore prevent increased migration of PHIIds to the lungs. Increase of PNAis in the lung after exposure to high levels of oxygen was associated with pulaonary edema and a leakage of senan albunen into the alveolar lumen. IPS significantly depressed both patblogies. Postulates that PANIs and PNAIs oontribute to response to oxygen toxicity by release of free Cii radicals. Used IIM50 as a scavenger to prevent damage. Works well but the severe garlic odor caused by DN6o would . make it an unattractive agent to treat patients. 7"hiourea (H2N-C:S-NH2) was then tried as a scavenger of CH radicals and found to be effective, but caused severe (fatal) pulrrot:ary edesa. Agparently the following chemical reaction oecvrs: OFI + ZU ~a CYnamide, which caused lung injury. Gynamide (H2N-C:N) is an oxidation product of thiourea. Separate stuc3ies daronstrated that the cyanavnide directly caused lung damage and a leakage of albumin into the alveolar spaoes. Dimethylthioura3 (DIm),(HfM-C:S-M-M ) was the next free radical sca- venger utilized. This aaTowid was nat oxidized ta3an amide and was quite effe~tive as a free radica]l scavenger. witbut causing lung dmnage. In futnre studies D1M will be the scavenger used in preference to DMSO.

-2- T«i: Cc~n -Wir. Tou'I'otsncco RssEnRCO-U.S.A.. INC. R. Harada: Dr. Harada has recently been appointed an Assistant Professor and has two years of support fz+o:n the Parker B. Francis Founc7ation. She has cbServEd mflamnatory changes in lungs exposed to hyperoxia, with a marked aocumualaticai of PMs. Studies with neutcopen; c rabbits exposed to hyperoxia stxywed a minimal d!eveloprert of lung injury. Concludes that hyperocia ney induce an increased release of chamtaxisLs frun PAFLs, which induoe migration of PMs to the lung. PAMs in culture and exposed to hype=cocia sivaaed an incr®ased xElease of LDH. FN evaluation of such PAMs showed ttem to have pynln:otic nuclei and waciaolated cyixD- pla4na. Such oxygen exposed (for 48 hyurs) damaged PAt4s were observed to show an in- creased release of a substance which uas daimtactic for PMNs. Seventy-two hour expo- sure was associated with a reduction in release of droenotaxina. Fr+esumably due to the severe damage to the PANIs by this time. o: gel separation, the dt®mtaxin appears as two camponents, ane large aaie small. The snaller appears to be peptide, the larger may be a protein. Dr. Harada's future plans include further separation of these dhe- nx"ctic substanoss, their purificatien and cha+acberization. Tte PAM also appear to release a factor which increased PiuYJ adherence to nylon fibexs. This factar also appears to be a peptide. M. BoMmian: An M.D./Ph.D. Assistant Professor in Pediatrics with a five year support f~ra the American Heart Association. Using a tissue culture moael, Dr. BcNman has obsexved that hype=;a injures eredathelial oells and that PMNs added to the culture then adhere to the injured cells (this might also explain the increased PNNs in the vessels of hyperoxic exposed luuigs).'t1K studies st:owad that the RSR of endothelial cells was essentially totally destr..yed by a forty-eight hour exposure to hyperoxia in culture, with a marked =elease of IDA by the damaged cells, in culture. Similar changes occurred in vivo. F+eels that such changes nay be responsible for the capillary leakage of albumin into the alveolar spaces off hyperoxic exposed lungs and may further act to accentuate the inflamnatoty response. Flvcther, the normal PMs utich adhere to the damaged endothelial cells may release proteases to oontribute to further lung da- mage. FITIURE PLANS: 1. Detecmine nechanisms of hyperoxia induced etxlathelial injury. 2. Evaluate response of Pl4iVs to erdo*he7ia7 cells injured by hypemxia. 3. Study the effect of adherent stasm:lated PWs an endothelial aells. 4. Study role of oxygen radicals in injury to endothelial oells (by hyperoxia or released fran PM+is). R. Tate: Dr. Tate is a seaartd year palirc+nary fellaMr who may join the faculty at the end of~)us fellowship. inbrks with Dr. J. Eieffnpx, J. Cannon and M. Shoemaker with an isolated perfused rabbit lung nodel. (See progress report) First observed that PMs stamulated by PAMs release toxic oxygen radicals wihich increase lung edsma and leakage of a]bmin fzcm capillaries. Unstimulated PMlis f:vn human granulanatous disease had no such effect .... appear to lack ability to Pro- duoe free oxygen radicals. Chmically produoed oxygen rad7rnlA (purine + xanthine oxi- dase) also produced lung edoa and leakage of albmin. Dam:age prevented by use of UNIIU as a radical scavenger. This group have undertaken a nunbex of other studies to show that oxygen radicals cause lung edam and alhamtir leakage whidh both appear to be linked to the presence of PAM activated P[Tls. Such PNNs may also have other effects, as in the production of athenosclerosis or in aqftseua. Concludes that oxygen radicals, either prodoaed dani.ca]1y or released by PAM activated PNNs play a major role in the path4genesis of puLronary disease.

, _3 _ __ THr COUNCIL FOR TOBACCO REBEARCH-U.•'l.a.. INC. FVl[JRE PLAffiNED n4VESTICMICKS: ~fS~~_~',((~lil Can scavengers block oxygen radical injury in Living anima].s? Can PMN injury be manipulated phannaoologica7ly, ie., with stet+oids? Iiow do other calls interact with oe]ls injured by Pms, e.g. , platelets? Can more specific indicators of endothelial danage than IDdi release and RM destruction be defined? Can serum factors or arachadonic acid metabolites whidz intaract with PM/OZ radical lung induced injury be identified? Dr. Repine, in the short tine be has been at NiaYab Warirtg, has asseabw an active enthusiastic group of young investigators around him who though irostly supported by fellowship funds fxnm othpx sources, are very acti.vely involved in this CTR project. Using various approadses, they are directing their activities to dp*p•++t;*+;++g to what degree and by what meahanisms free xadica]s of axygen, such as nay be Modu~ced by PAM activated PMs, oontritxrte to pulamnaty edeea, albianin leakage froan capillaries into alveolar sacs, P*+dothel; al in jury (lriay aontribute to a].bmin leakage, but may also play a role in athetosclerosis) and a possible oontribution by activated Prgis to ee- physema by release of p=otease. Their progress since Dr. Repine joined webb Waring (antedates the start of the grant by about a year) has been considarable and at this point merits aontirnaed support. D. H. FORD R. C. HOCKE!T /am