Biological and Medical Research Group (H-4) of the LASL Health Division, La-3848-MS (July 1966 Through June 1967)

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LOS hUM0S SCIENTIFIC LABORATORY University of California LOS ALAMOS0 NEW MEXICO Biological and Medical Research Group (H-4)- of the Health Division-Annual Report July 1966 through June 1967

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This LA.. .MS report presents the &&us of biological and medical re88aFch in Group H-4 at LASL. Previous A n n d Reports in this series, all unclassified, are: This report, like aher special-purpose documents in the LA.. .MS series, has nc4 been reviewed or verified for accuracy in the interest of prompt distribution. Printed in the United States of America. Available from Clearinghouse for Federal Scientific and Technical Information National Bureau of Standards, U. S. Department of Commerce Price: Printed Copy $3.00; Microfiche $0.65

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10s ALAMOS SCIENTIFIC LABORATORY University of California LOS ALAMOS NEW MEXICO Report distributed: January 23, 1968 Biological and Medical Research Group (H=4) of the Health Division-Annual Report July 1966 through June 1967

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Abatraak of Cel lulu Radiobiology Section Publiaations and Manuscrip+s Submitted 11 S thul at iam of Amino Acid Incorporation by CoPp.tence-Stimul&ting Factor from Hemophilus influonrae 11 2. Stimulation of Competence of Hamphilum influema. by a C e l l Free Pactor(s1 3. Eleatroaie Effects i n Horse Liver Alcohol Dahydrog.na8e Catal yr it 12 4. Prepuatioa and Assay of PhoBphorylating Submitochondrial Particles: P u - ticla8 from Rat Liver Prepared by Drastic Sonication 12 5. Polyriboadutylate Synthesis by Nuclei from Developing Sea Urchin E ~ lyos. I. Characterization o f the ATP Polyurase Reaction 12 6 . Polyriboadenylate Synthesis by Nuclei frcm Developing Sea Urchin Embryos. 11. Polyriboadenylate Priming of ATP Poly m rue 12 7. Configuration Change of Surface Si ali c Acid during a to a is 12 8. Requrer&tion of Si ali c Acid on the Surface of Chinese Hamstar Cells in Culture. 1. General Characteristics of the ReplaceKent Proces8 13 9. Rmguruation of Si ali c Acid on the Surface of Chinese Hamster Cells in Culture. 11. Characteristics of Glucosuaine Incorporation 13 10. RNA Stability and Prowin SynthSiS i n Relation tothe Division of M-- 11. Properties of Mitotic Cells Obtained by Shaking 13 12. L i f e Cycle Analyois of N.nrmaliWh Cells. 111. The Inhibition of Division of Chinese Hamster Cella by Puromycin urd Actinomycin 1 4

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Owrnic Sc in dll at ioa Detectors md Their Uuin the StudY of Body Carpom it ion Differential Rasgonse of Two Derivative. of the Bm21 1I.aut.r Cell to Cell Growth and Division. I. Mathematical Analysis of C e l l Volume 15spectra Cell Growth and Division. 11. EXpuiP.ata1 Study of Cell Voluma Sp.c+ra 15 Effects o f uaoarplers of oxidative Phosphorylation on the Iafoction of StruCtUXal Cb.nqu lurociatmd with Acid Xaactiwtion of Home Liver Alcohol 16Conjugation in Stentor coeruleus 16Studies of Surface Glycopeptides of Cells in Sumpamion Culturm Evolution of Enzyme mc at fon al Spocificity: Hod.ls i n Microbial Genetics 17 The Effect of Thymidine on the Duration of C1 in the L i f e Cycle of Chi-mae PrOP U ties of Utotic CelU Ptep8red by m chi oic ally Shrking Monolayer 1 7Bioch.laic.1 &lance and Synchronized Cell Cultures Effmctr of Detergent on the Structure of Horse Liver Alcohol W h y & o g . a ~ ~ c 18

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31. E8sential Eioaynthatic Activity i n Synahronized -1i.a Cells 19 f o r UWOUl RNA in GI urd G2 19 33. SU &ad Protein Synth u is in Relation to Oral R.g.ner8tion in the Ciliata CIUPTER 3 - MOLECULAR RADIOBIOLOGY SECTION 2 1 after Diphemylamina-Formic Acid Degradation 28 4. Urh - Deoxycytidlne following Chronic Whole-Body frradiation of thm 6. Amino Acid Analyris at the Nuro ~ aol eLevel 4 1 8. C huac teri ration of Chinese Hamter ovuy Hiatone FrAction8 49 9. The Synthelis of Histone Fraction8 during the L i f e Cycle of Marmo.liur 10. Synchronization of Chinese Hamster Ova ty Cells in a Tisrua CUlt UZ8Pilot

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Abstract. of Mlecular Radiobiology Section Publiacrtions and IUnwcripts Suhr it k d 59 59i. D 1 8 c Electrophoresir of Bovine Thynrus Histoner 2. Nuclear Histonma urd Early anbryogOXIe8i8 of the Chick 59 3 . Inoorporation Efficiency of Small Olig0-5'-Nucl.otid. Initiators i n the Teminal Deoxyribonuclootide T rur sfu & s e Reaction 59 4 . Syntherir of Polyad.nylate:Polyuridylak Catalyzed by Ribonuclaic Acid Polymerase i n the Absence of a Tamplate 59 5. Heteropolynuclootid. Synthesis with Terminal Deoxyribonuclootidyltrms6.Purification and Properties of Ribonucleic Acid Polymerase from Escherichia 7. Synthesis of a Polydeoxyribonucleotide Containing an Internal Pyroghooph8tO 8 . Enzymatic Synthesis of a Three-Section Block Copolymer of Th pid y late, 9. Limited Enzymatic Addition of 0.oxyribonucleotide U n i t s onto ChuPic8liy 10. Tamplate-Induced Dissociation o f RNA Polprase 6 1 11. The Intra call u lar Distribution of Basic Proteins i n the Chinese Haamtor CHAPTER 4 - MUMALIhN UADIOBIOLQGY SECTION 6 2 Expomure aa a Function of Age i n Mice 6 2 642. Effective Residual Dose Studies on Monkeys and Dogs

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4. Oba uva th m on Progeny from C o ~ e cut iv eGaneratione of X-Irradiated Ab8traat8 of Yl nrl i a a mdiobiology section ~ublicatioas and Manu c tip t. Submitted Col pu at ivo ~crpopulationRecovery of Circulating EXythrOWte@ following Graded Secolld Gllu-Ray Exposure8 i n Hico A Spontaneous Hereditary Mutation for a Recurrant Hairless Condition i n R8diobiological Factors in Manned Space Flight Survival Thin nico The Effmctive Rosidual Dose Concept of Radiation Injury -- 1\ Test with 1. Cee iw l37 Cantent of New Mexico Control Subjects: July 1966 through 2. Protraatd Peoding of Plutoniua-23E Microsphmru t o Bmgles. I. Macroand Micro8copic Observations of the G8strointertinal Tract and Estim ~ tas of Deposition in Liver, Splem, and Lymph Nodes 3. Long-Term Retention of Cobalt-60 by Rats for: Cesium-137 Body Burdurs i n Nan Effect of Pilocarpine Nitrate on the Excretion of Strontium-05 6. A Radiation Protection Guidm for Silver-110 Es tim ted from Retention i n

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941. Comparative mtrbolisa of R.dionuc1id.s in m s . 111. Retention of 942. Larg-Tera i n vivo Retention of Cerium-144 by Boagles 943. Large Organic Scintillation Iktectors 4. Wuourt of Discrete Particles i n the 100- t o 20Q-nicron Dim0t.r RU1Qe through t h e Gastrointertinal Tract of Laboratory Animals and W.n 9 4 6. Accolerating tho Turnover of Intunally-Deposited Radiocm8im 95 957. Clinical Uses of Whole-Body Counting 958. Cesium-137 Body Burdons i n Wm: January 1965 t o Doc.aJ>ar 1966 CHAPTER 6 - BIOPHYSICS SECTION 97 971. ~n Auto antic Cell Cowtor 2. The PlWtemcmnt Cell Photaarter: of Biological Cells Stained with Fluorescent Dyes Abrtracts of Biophyricr Section Publications and Manuscripts Submitted 1. Vol w Distribution and Separation of Normal 8umn Leucocyte8 2. Intercompuison of the Reliability of Body 13’Cs Moa sur r ~ .ntr on Hurvn

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1ish.d or sub rit tod for publicatrcol ha8 not b a u ~rrwrittmn. Abatractr of these journal tute mst hportmt puts of this report. t.ko advantage of t h e Laboratory's uniquo faciliti.8 urd taleat8 urd which are of funduaontal scientific rig n if i cm ce in radi ation biology. The rtaff and fbCiliti.8 out dolay progrumatic problem a8 they m y ariro within the Laboratory o r olsewhere. Over tho part fw yaur incroaaed emp hu is ham bo- plicod OIL studies a t the moleculu sults froar tho.. new pro j ecw arm appoaring at M increasing rate. During the next ye u met of these projoata will oontinue with the total group effort raining at about tho prasent lmvol. Connidoration will bo givon to initial rpeciffc planning of facilities and experiment. f o r the proposed bi-dical research addio logical urd Modical Rorebrch Group €or the puiod July 1965 through June 1966 appoared Tho provioru annual rmport of the Bi-

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WBLICATIOMS AND ABSTRACTS OF MANUSCRIPTS SUBMX'ITED STIUULATION OP AMINO ACID INCORPORATION BY Cox. Bacteriol. Proc., Abstract No. GP-29 Whmn cultures of Hapophilus influenzae ability) are re8USp.nd.d i n sterile fil - trato8 taken from cultures of highly compc- +ant cells as compared t o rasuspansion i n fruh diu, the forrur bacteria are stim - u late d to develop competence with a shorter initial delay period and a farter rate once developmont starts, and 25 t o 30 fold more cornpatant cells are attained a t mwillllll~ t o determine the mechanism of action of Studies of incorporation of 14C-labaled amino acids into TCA-precipitable fractions as part ic acid are incorporated at different rates i n the pruence and absence of CSfactor. The shapes of iaotope incorporation curve8 follow closely those describing stimulation or lack of stimulation of development of compatmcc. Although the last two of tho aforementioned amino acids and first two of thoae mentioned below are reported t o be essential for development of a reaoonablo level of competence i n A dafined medium, their incorporation rates are d i s - Work is currently i n progress was not stimulated by CS-factor. results were discu8.o.d relative to the physiological state of Competence. a tracts, IInd International Biophysics Congrws, Vienna, Auatria (19661, Vet lag der Wiener Medirinircbul Akadde, Ab8Uact Recent reporta doscribe control i n reveral bacterial system of dovelopunt of competence (i.e., t h e physiological state neceaaary for tram form tion) by out-lecular matabolic products of cqunt cellr. Several repor-, however, have pteeurted data showing that no such control 8 y e t u could be detected with 2. influonzae. A systematic reappraisal of the lattmr tran8formation system has reveAled that an inter. cellular control of competence development can indeed be dw nat rated. The control 10 mediated by a factor, or factors, , ~ tl out one of which is dialyzable, and preparation tence are resuspended i n sterile sup.rnatants taken from cultures of highly competent cells aa colapared t o those takea from cultures of low-level competence o r fresh media, the former bacteria are stimulated t o develop competence with a shorter initial delay period, a faster rate once development starts, and a total of 25 to 3 0 fold more competent cells at maximum development. Stimulation of competence is greatly reduced by treating the coll r with puromycin and by reducing t!!o cell incubatured bacteria from the early to mid-log

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phases of grarth aro aost sonsitivo t o rtimulation. Progrerr on pur if i ation and identif i cation of tho factor(s1 and infonuation on its ~ chan ir mof action weto dircussod. ELECTRONIC EFFECTS I N BORSE LIVSR ALCOHOL cohol dohydrogoaarccat8lysod r duct ion Of tuted benzaldehyde compound8 have been made. There w a s no apparont doviation from the ordorod aechurirm port u late d for the reaction with myof the a ub stratos oxamind. The values of the kinetic constants @oand aldehydo substrate. Tho values of 1/0, increased with tho in ere wing alectronwithdrawing paror (0 valuo) of the p.rlsubstituent. Tha r orul t sue conristent with tho postulated reaction mechanism involving tho tru rr fer of a hydride ion toor from m alcoholate ion intermediate. were indopendent of tho nature of tho PREPARATION MID ASSAY OF PHOSPHORYLATING SUBMITOCHONDRIAL PARTIcLEst PARTICLES PROM RAT LIVER PREPARED BY -TIC SONICATION, Abstractod i n Lon AI-s Scientific POLYRIBOADENYLATE SYNTmSfS BY NUCLEI FROM ACTERIZATION OF TIiE ATP POLYMERASE REACTION, from dovoloping soa urchin d r y 0 8 havo boon UTP, and GTP inhibit A18 incorporation and are n o t polyurired. Dmoxyadononinc triphosphate is incorporated approximately onetenth as efficiently as AHP. pyrophosphate, but not phosphate, inhibit8 POLYRIBOADENY~~SYNTHESIS BY NUCLEI FROM DEVELOPING SEA URCHXN EMBRYOS. 11. POLYRIEOADENYLATE PRIMING OF ATP POLYMERASE, An ATP poly w rue present i n nucloar shown t o bo primod by low molecular-weight polytiboadenylic acid, low molecular-weight yeast RNA, and sea urchin RNA. high molecular-woight poly Awas activatod by troatanent with hog liver nuclear., which cleaver t h e polymer leaving a froe 3 ' - hydroxyl group. Treat m nts that cleave tho polymer having 3'-phosphate groupo did not activate poly A. Priming activity of poly A was inhibited by dATP. The acid-insoluble product of the poly A-prinwd reaction wntahcrd an average of 3to4 AMP rosidues. Th ue was no evidence that chains could be initiated by tho polymeraso, a s no [5s-14C]-A)IP was recoverod from an alkaline hydrolysate. CONFIGURATION CWGE OF SURFACE SIALIC ACID Abstractod i n Loc A18mos Scientific The char actor is tics of an ATP polymerase which occurs i n nucloar preparations

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REGENBRATION O? SIALIC ACID ON THE SUR?ACE O? CXINBSS HAMSTER CEUG IN CULTURE. IAbstracted in Loa Ahma Scimntific REGENERATIO# O? SIALXC ACID ON THE SURFACE O? CHINESE HAMSTER CEUS IN CULTURE. 11. CHARACTERISTICS OF GLUCOSAMINE INCORPORACulturod Chinose hamstmr cells incorporated radioaotivity from glucos.mine-114C into surface sialic acid and into t o medium containing unlabeled glucosamine progressively lost counts tothe nwdium for mrny hours. Such chasm experimsnts suggeated a mtm rapid turnover of trypsin-rmovable matdrial than of surface-bound a i ali c acid. Further studims ofthe reganeration of surface sialic acid showed that the actinomycin D-resistant portion of t h e procmas involved emergence of an intracellular precursor onto the cell surface. hn earlier portion of the process was inhibited by actinomycin D, and a t least three step8 were inhibited by puromycin o r cyclohexRNA STABILITY A#D PROTEIN SYNTHESIS IN RELATION TO TIiE DIVISION OF MAElMALIAN CELLS, sen. Proc. Natl. Acad. Sci. 56, 1520-1527 Experiamntal determinations of the stability of messengar RNA (m-RNA) provide a useful parameter to aid in understanding the mechanism of transcription and translation of gonotic infomation into fuactioaobtainod by a m8thod i n which rpmcificity is obt8h.d by observing a uniqum cell function (in this case, cell division) and i n which highly precime timing is attained by uam of tho luthods of life cycle analy8is. A prtten of drug inhibition is chosen t o produce a populatian of pr d to tic call8 which have afull colplaaurt of m-RNA's but not o f the protmins necmssary for diVi8iOn. Tho calls arm held i n this 8tate with both protoin and RNA synthm8is inhibited for increasing p c rid8 of time to allow loan of masrengers t o occur. They are than relea8.d intoa madfur in which protein synthesis is permitted but RNA synthesis is still blocked. Thm fraction of cells able t o complete mitosi8 under these conditions is taken t o be a meaaurm of survival of m-RNA through the period of pret the experimental data, w e find th&t the mean lifetime of m-RNA coding for protein msrontial for division of the Chinese hamster ovary cell is approximately 2.9 hours. Ebraloying this assumption t o fntmrPROPERTIES OF MITOTIC CELLS A quantitative study of propertie8 of rynchronirmd Chinese hamster cells prmpared by selective shaking [Robbins and Marcus, Scimnce 144, 1152 ( 1 9 6 4 1 1 ha8 been nuda. Mitotic cells were separated by mechanical shaking and imawdiately placed i n SUSp8ASiOn culture. Growth rate and differential miqu at intervals thereafter to detmrminm the timing of major events of the mitotic period. Populations were routinely obtained i n which X. I. dropped from 0.95 t o 0.01 in 15 minutes. Chilling immediately after ress. Subsequent resuspension i …