Tobacco Documents Online

:.: rPHYSTOI,OGY OF THE RESPIRATORY SYSTEhI: THB COUNCIL FOR TOBACCO RESriARCH - U. S.A. 633 TIICFtI7 .dVENLTE NMSr YOR%, N. Y. 10017 Date: ? August • .' Name of Inve3figatar(sj: (iric(ude Title and Degrees) • :..; Theodore K. F i n 1 ejr;-'M,D. , Assoc i ate Profes sor of Med i c i ne -'Aaron J. Ladman, Ph.D., Professor and Cha•irman, Department of Anatomy. 2. Institution & : Address: : r e 2'ic' : ' The Undvers_ity_ o• N6r Flex i co School of Med i ci ne -915 Stanford Avenue N.E. Albuquerque, New Mexico 87106 3: Short Title of Project:: .. . . -Effect of cigarette smoking on lipids and morphology of alveolar lining material AS and macrophages. Proposed Starting Date: ~ ~' January 1, 1968 :: 5. Anticipated Duration of this Specific Study; ,. 3 years to December 31, 1970 '' 6 Brief Descripton of. Objectives or Specific Aims: . Four studies will be undertaken: :' d a d r h li l "' f ff th i h f i ki i D ~ ~ s n mo o n on e o ono e e ect o c nat t eterm 9arette smo (a) 9 P P 9Y:w;~ . non-smoking humans. of alveolar lining material and macrophages recovered by saline lavage of smoking and K. ~ lo `' '`Determinetion of acute an b d chronic effects of smoking orn liPi. ds and morPho { ),._._..._ 9Y of dog alveolar"lining material and macrophages in situ as well as on fractions recovPred ; a 1i i ne lMage. circulating whi!te cells with radioactive lipids and amino acids and monitoring!the transport of these labelled'cells in:to the alveoli of the lung by radioautographic microscopy. ..(c) Determination of the effect of cigarette smoking on the incorporation of radioactive lipids into alveolar lining material and macrophages of the dog. (d)"~ Determination of the origin of the alveolar macrophage by labelling ,7. Give a Brief Statement of your Working Hypothesis: -.. . . ,- - . . . (See attached sheet) _ ~_.~,. . .....-.~........ ......... ~w+ir:y.~,,.._. ......:..,a::u.,...~:.........~.w~...~..1.~..:...~..

Detaili of Experimentol Design and Procedures: (Attach Separate P4ges) 9. Physical FacSlties Available (Where Other than Administering Organization Indicate Geographical!tacalion) See Attached Pages ( 10. Additional Aequiremenfs: See Attached Pages C 11. Blographical sleetches of all princlpal and professional personne( (qppend) See AttacHed Pagt?s I 12. List of publications: (Five most recent as pertinent) (append) tSee Attached Pages 17"ft. -NUM.- E'-"-PMNWM Eft"ft-=,F-

;; Preliminary experiments have shown morphologic differences in* the alveolar macrophages recovered by saline lavage of smokers when compared to non-smokers.'~.;" The alveolar macrophages of smokers contain an increased number of inclusion " bodies of various types. In addition, the fatty acid distributi-on patterns.of the phospholipids extracted from both the cellular and non-cel,lular fractions of the lavage differ in smokers and non-smokers. There appear to be more unsaturated long chain fatty acids in the phospholipids of smokers. More studies. are necessary to confirm these.observations. . _ : . . • . .. . . ,.;: The incorporation of labelled lipids and thei!r precursors in alveolar` macrophages of control dogs and; dogs exposed to cigarette smoke will be studied to determine aitered: l,ipid pathways. We feel the above studies are necessary before more detailed studies of alterations in lung morphology and lipid metabol- EXPERIMENTAL EXPERIMENTAL DESIGN AND PROCEDURES: Surface activity of the acellular alveolar tining, material will be measured on a modified WilheTmy balance. ism in patients with various lung diseases are undertaken. In addition, studies to determine the origin of the alveolar macrophage will be attempted because smoking may alter not only the quality but also the quantity of these cells. Lipids will be extracted from the cellular and acellular alveolar lining material by several Folch washes using CHC13•:Me0H:H2O. The various classes of _ --••lipids (neutral and phosphol i pilds) will be separated by thin layer chromatography using silica gel H and CHC1 :MeOH:acetic acid for phospholi.pids and petroleum .ether:diethyl ether:acetic Kiid for the neutral lipids. The lipids will be local'ized on the thin. layer plates with 2,7 dich°lorofliuorescein, and removed by ; scraping. Fatty acid methyl-esters will be formed,by refluxing!in Me0F1 and H SO and will then be separated by gas liquid chromatography at 210° using a butanedtio ': succinate column. : The lavage sediment containing alveollar lining!material and cells will be centrifuged at 10,000 x G at 0° C for 20 minutes, fixed in cold,2.5% glutaralde- hyde buffered with 0.15 M Na cacodylate to pH 7.2 for two hours, washed:several times in•buffer, fixed in cold phospha.te buffered 1% 0s04 for two hours, rap-idly dehydrated in ethanol and then embedded in Epon 812. Sections for light and electron microscopy will be made from the eporn blocks. In attempts.So isolate relatively pure monocytes from circulating blood, buffy coats from dog bliood obtained by exsanguinati:on will be processed by an ~ electronic separator so that monocytes can be separated. They w-H-l be labelled with rad.ioactilve:substances and injected into dogs. Their course into the alveolar spaces will be followedd by appropriate radioautographic microscopy. ~ The exper i,men-ta l des iign w i 1 l emp 1 oy the procedures ou t l i ned i n the f i rs t three paragraphs above, on humans who are smokers; non-smokers will serve as controls. (The appended reprints contain further information about our methods ~.:•. t..: and observations and are included as an aide for the reviewer)

9. PHYSICAL FACILITIES AVAILABLE `Tr has approximately 4,500.square feet of research~.` The Department of Anatomy. . ,_. ..,space which is available for use_in support of the outlined program. This includes`~ •three electron microscopes, four ultrami-crotomes, two fully equipped photographic' , darkrooms with the supporting equipment to do excellent photomicrography.' Suitable 'supporting technical and secretarial personnel are currently employed ~ :.to assstn tempemenaton•o te program~ ..: ;.. • .. - .. . . . '- . . •_ _ . ", .. . .~• _ , . . ,- . , .. . ..A ' outlined. has the equipment and facilities to accomplish the biochemical procedures as '. r, medical complex. Dr. Finley's laboratory ds composed of 1000 square feet and,:;~= Members of the Department of Medicine have laboratory areas within'the :f Sufficient animal quarters are present to provide adequate space to house and ma i nta i n the needed suppl i es of dogs. catheter. . hospitals, to perform the lung lavagos on human subjects. X-ray and fluoros- copic equipment are at hand to monitor the positionirig of the intraalveolar Albuquerque Veteran's Administration Hospital, the Universitiy's teaching Rooms are available at the Bernaiil'llo-County-Indian Hns.pital; and 10. ADDITIONAL REQUIREMENTS that a base-line for the non-smoker and the smoker can be establi!shed. So far, ';` We are attempting to survey a sufficiently large number of humans so ,,., $25.00 to each individual who has volunteered for the lung lavage procedure. y g n e av g p p ,, v a ne n v der aic! er fift subjects have u on ast we ha s li a e l the e o ] d a iil rt k the th d h I t t ti ld th it l h e e e even an un ow reac a w un a n r on s ou occur, osp r e ma i ntenance and care of the pat i ent. Add i t i ona 1 funds to cover such added costs should be budgeted so that effective patient-care can be provided; . . .. :*,: . i

t CURRICULUM VITAE P-1 ' ,.Seattle, Washington Assistant Resident in Medicine University of Ca!liifornia Medical Center, 1955-1956 San.Francisco County Hospi,tat, 1956-1957- r POSITIONS: Assistant Resident in Medicine Research Fellow, 1957-1958 University of Buffalo Department of Physiology CI i n ica I Instructor i n Med ic i ne, 958-196.f Cardiovascular Research Institut+e= Unavers it~y-of Ca I' i forn i a Med;ica II Center Associate Staff Member, 1960-1961 Cardiovascu;lar Research .Institute University of California bedical, Center ~ • SOCIETIES: R: REDACTED MATERIAL University of Washingtori;: IS44-1950! Degree: B. S.. (Chemistry) - Johns Hopkiins Medicai Schoo-l-, 19504954 Degree: M. D. San Francisco County HosRital, 1954-1955 American Trudeau Society Research Fellow, 1958-1961 Associiate Professor, July 1964 and B'iiophysics; Director of Anesthesiology Research University of Washington School of Medicine 1961; Assistant Professor of Anesthesiology, Physi:o,logy Assoc iiate Prof essor of M_ed.Ic i ne, Un ivers i ty of uew-Mexico School of Medicine., September 1964 to• present. American Physiological Society We.stern Society for Clinicali Research American Society of Clini.cail Investigation Amer ican Fed'erat ion for CI i ndca I Research

B1BL10GRAPHY - T. N. Firi:ley, M. D. Nbrgan, Thomas E., T. N. Finley, Gary L. Huber, and Helen F-ialkow. -Alt+erations in Pulmonary Surface Active Lipids duciing Exposure to • Acute effects of saline lavage on pulmonary mechanics and morphology. J. Clin. Invest. (irn Press) Lung Lipids in the dog. Bioch!im. Biophys. Acta, t06-:403-413,1965. 3.A Huber, G. L., Edmunds, L. H. and Finley, T. N. of the Composition and Surface Activity of "AJveolar11 and Whole 2. Morgan, Thomas E., Theodore N;. Finley and Helen Fialkow. Comparison Increased Oxygen Tension. J. Clin. Invest. 44:-1737,II965. . Finley, T. N., Swenson, E. W., Curran, W. S., Hu,ber, G. L. and Ladman•, A. J. Bronchopulmonary lavage in normal subjects and patitents with obstrucfiive lung disease. - Ann. I n-tt . Med. 6b : 65 l, 1967 . 5. Finley, T. N., Ladman, A. J., Brewer, L., andi MrKay, ML B. A morphological and' 1 ipid ana,lysis of the alveolar I ining, rreterial in the dog. J. Lip-id Res. Un Press)

R: REDACTED MATERIAL CURRICULUM V'fTAE i MAJOAt' RESEARCHI .:1NTEREST: -Aaron J: Ladmany Ph. D. P-1 :" Jama ica, New• York, U. S. R: New York Univers•ity, New Yor:k, N. Y. Indiana University, Bloomington, Ind. American Cancer Soc., Postdoctoral Research Fellow 1952-1955; Special Research Fellow ofs U.S.P.H.S. 1955-1957; Career Development Award 1962-64; Visiti.ng.Summer Investi.gator, Jackson Laboratory, Bar Harbor, Maine 1949-1951, 1953•, 1954, t956; - 1957, 1959; Fellow•of American Associ-ation for the Advancement of Sc ience, 1966.. Funct iona I Cytology-aind Cytocfiemi stry RESEARCH AND PROFESSIONAL ..EXPERIENCE: Research Assistant, Jackson Laboratory, Bar Harbor Maine, 1947-1948; Teac h i ng!Fe I I ow im-Anatomy, Un i vers i ty of C i nc i-nnat i, Depa rtment of Anatomy, C i nc i ranat i, Oh i o, 1949-1949; Teaching Fellow i:n Anatomy, I.ndiana Uni.versity Department of Anatomy, Bloomi-ragton, Indiana, 1949-(952; Research Fel low im, Anatomy,' Harvard Medical School, Department of Anatomy, Boston, Vass., 1952-1955; Associate-in Anatomy., Harvard Medical School, Department of Anatomy, Boston, Ma,ss., L955-196D; 1964 to presen-h. ; e ca ennessee - n ts Memph i s ,.,, Professor of Anatomy, Chairman, Department of Anatomy, The Un ivers i ty- of New Mex ico Schoo I cf Med;i c i.ne, y, 1961 I 964 d i I U T M i Un ivers.ity of Tennessee" Assoc iate Professor of Anatom

. ~ B1BL:OGRAPHY - A. J. LADMAN, Ph.D. .: Ladman, A.J. and F i n iey, '1'. N. E1 ectron m1croscop-i c observat i ons o pulmonary surfactant and the cells which produce it. 2. Finl•ey, T.N., Swenson, E.W,., Curran, W.S., Huber, G.L. and Ladman, A..J. Bronchopulmonary Lavage in Normal Subjects and Patients with Obstructive Anat. Rec. 154:372,1966. (Abstract) .Lung-Disease. Ann. Int. Med. 66:651,1967. Ladman, A.J. The fine structure of the ductuli efferentes of the opossum. Exfoliative cytology of the lung alveolus: Preliminary electron microscopic observations on cells obtained i_n,vivo from human lungs. Elec. Micr. Soc. Am. 1967. (tn Press) Anat. Rec. 157:559,1,967. Finley, T.N•., Conley, G.R., Huber, G.L. and Ladman, A.J. of human alveolar lung,mate'ri!al and cells. Clin. Res. 14:365,1966. ,(Abstract) Ladman, A.J., Pratt, S.A., and Finley, T.N.

R: REDACTED MATERIAL Chemicals Glassware Reagents EM,supplies L Consumable Supplies (list by cntegories) C C. Other Expenses (itemize) Dr. Susan A. Pratt - electron microscopist Technical Laboratory Technician: Secretarial assistance Animals, maintenance & procurement 1/2 service contract for EM• Consullting fee to• Dr. Swenson Patient; reimbursements 100. 100 25• Sub-Total -- .. : - --7711 Amount 500- .. 500 --- 500 1,000 Sub-Total $,2,500 1,000 700 500 3,000 Sub-rotalt D. Permanent Equipment (temize) I E. Overhead (15% of A+B+C) Estimoted huture Requiremenls: Year 2 Year 3 s,200- $3,405 Total $26, f05 Salaries Consumable Suppl`. Other Expenses Permanent;Equip- _ Overhead ~ Total P- 2,500 4,200 P-. - 2,500 4,200 11 is understood thotAhe opplicant and institutional officers fn•applying for a grant have read and found acceptable the Council's "Stotement ofi Policy Containing Conditions andTerms Under Which Project Grants AreMade."' 3,368 25,818: /T 4 26 ~ Sgnature Signoture Telephone ~-r-i ewn.,, orr ..r tn. t.,rk.ttoe Telephone C GC!M

I 4. Other Sources of Financial Support List financial support for re.earch from all cources, including own in.titution, for this and/or related research projmcts. Current Titie of Project Fine structure of tissues during transport behavior ibid. Respiratory Distress - Role of Lung 8 Blood Lipids National Sourc. Institutes of Health Amount $38,435 9= Durotion 1-66- Grant #1 R01 GM14435-01 8-31-67 ibid. Grant #1 RO1 GM14435-02 $28,676 9- 1-67- 8-31-68 National Institutes of Health $28,500 9- 1-67- Grant #5 ROl HE09491-04 8-31-68 S t w