Biological and Medical Research Group (H-4) of the Health Division, Annual Report, La-3132-MS (July 1963 - June 1964) (Redacted)

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LOS ALAMOS SCIENTIFIC LABORATORY OF THE UNIVERSITY OF CALIFORNIA LOS ALAMOS NEW MEXICO REPORT WRITTEN: July 1964 REPORT DISTRIBUTED September 14, 1964 BIOLOGICAL AND MEDICAL RESEARCH GROUP (Si-4) OF THE HEALTH DTYISION -- ANNUAL REPORT JULY 1963 THROUGH JUNE 1964 Contract W-7405-ENC. 36 with the U. 9. Atomic Energy Commission Au U...US mporl. am Informal doaumda. u a W y pmpued lor a apcial purpore and primully prcparad for UM wtthtn t b Labontory rather than for general dlbrlM1m. Thlr report h a not ben edlted. rodewed. or ver If Id for accuracy. All LA.. ,MS reporta express tho Wwr of the authors am of the t h e they were written and do not accearorily reUect the cptntonr of th* Loa Alamos Sdentlflc Laboratory or the flnal oplnion of the authors on thr subject.

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Ruthenium-106 i n Mice, Wte, and Dogs: Inter specific Comparisons The Retention of Manganese-54 by Mice, Rats, Dogs, and Monkeys The Effect of Temperature on the RetenThe Lack of Effect of Water Loading on the Retention of Cesium-137 by Rats 37 Simultaneous Jdeasuremont of Gnmma-Uitting Radionuclides by Liquid Scintillation

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6.Effect of Specific Activity on Stable Studies withCemium Turnover by Man: 7. Effective Retention of Parent e rally by Mice: Comparison of Intra peritoneal and Intravenous Administrations 8. Whole-Body Retention of Orally and 9. Lo8 Alamos Small Animal Counter: LASAC 111 76 10. Comparative Yetrbolic Data f o r Macaca Mulatta and Macaca Speciosa Monkeys 84 11. Retention of Silver-110 by Rat6 after Oral, Intra peritoneal, and Intravenous AclministraAbstracts of Mammalian Metabolism Section Publications and Manuscripts Submitted 100 1. Effect of Environmental Temperatures on Retention of Cesium-137 by Mice 100

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2. Electronic Processing and Analysis of 3. Metaboli8m of Radiocesium by Man and Selected 4. Distribution of Cesium-137 after Chronic Exposure i n Dogs and Mice 5. The Effect of Environmental Temperature on Retention of Chronically Administered 6. Application of Regression Analysis tothe CHAPTER 3 - MAMMALIAN RADIOBIOLWY SECTION 1. Veterinary Medicine and Surgery in t h e Biological and Medical Rese8rch Group Soctlon 1 - Breeding Characterletice of - Mice from 5 , 10, 15, and 20 Generations of

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Section2 - Radiation Son m lt iv it y in Mice from 25 Genmrationa of X-Irradiated Siren J. F. Sprlding, 0. 9. Johnson, and 5 , 10, and 15 Generations of X-Irradiated of Irradiated and Noma1 Ancestry t o a Nonand Clinical Blood Chemistry of Mice of Studien on Hewglobins of Mice of the an Index of Activity of Mice of Irradiated and Normal Ancestry Abstracts of Mammalian Radiobiology Section Publications and Manuscripts Submitted 138 1. Genetic Effects of X - Irradiation of lo, 15, and 20 Generations of Male Mice 138

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2. Longevity of First and Second Generation Offspring from Male Mice Exposed to Fission 3. Comparative Fundamental Physiological Parameters of Macaca Mulatta and a New Laboratory Primate, Macaca Speciosa 139 4. Dose Rate Effects on Lethal it y of Mice Exposed to Fission Neutrons 5.Physiological Capacitance: A Study in Physiological Age Determination 6. Dose Rate Effect on Survival of Mice during 7 . On the Question of Cyclic Variations in 8. The Aboence of Avoidance Recognition by t h e Mouse Subjected t o Chronic Invariant Gamma-

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CHAPTER 4 - BIOPHYSICS SECTION HUHCO 11: Status Report Contamination of Materials of ConstrucCeslw-137 Body Burdens of Control Cesium Calibration Committee Data Analysis by Computer Electronic Cell Sizing. I. Effect of Electronic Cell Sizing. 11. Influence of Physical Phenomena on Meamred Spectra 190 Electronic Cell Sizing. 111. Erythrocytes 197

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Abstracts of Biophysics Section Publications and 1. Radioiodine Metabolism i n Children and Adults after the Ingestion of Very Small 2. Computer Techniques i n y Spectrometry 3. On the Retention of Cesium-137 i n People 5 . Large Organic Scintillation Detectors CHAPTER 5 - CELLULAR RADIOBIOLOGY SECTION 1. The Effect of Actinomycin D on Premftotic 2. Replication of Mengovirus i n Chinese Hamster 3. AS table Protein Fraction Associated with the Mitotic Apparatus 218 4. The Energy Metabolism of Synchronized Cells 224

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Abstracts of Cellular Radiobiology Section Publications and Manuscripts Submitted 229 Culture and Slide Preparation of Leukocytes from Peripheral Blood 229 Synchronized Mammalian Cells in Suspension Life Cycle Analysis of Yp11'1Ian Cella. Cells from the Chinese'Hamster Ovary Grown in Suspension Culture A Digitized Comprrator for Karyotype Anal - Qur nti ty FWoduction of Synchronized Mammalian Cells in Suspension Culture 231 Synchronized Mammalian Cells : An Improved Method of Study by Electronic Cell Counting 23 1 Synchronized Mammalian Cello: An Experimental T e 8 t of a Model for Synchrony Decay 231 Culture and Slide Preparation of Leukocytes from Blood of Macaca 232

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CHAPTER 6 - MOLECULAR RADIOBIOLOGY SECTION Nuclear Magnetic Resonance Studies of Pur if i cat fon of DNA Polymerase from Purification of RNA Polymerase from nucleotides to Their 5'-Triphosphates 242 GRADPRO:! A n Equilibrium Density Gradient Ultra centrifugal Computer Program Spectrophotometric Analysis of Nucleic Chemical Polymerization of aTri nucleotide 252 Chemical Studies on Nucleotide Polymeriza-

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10. Synthesis of Labeled Deoxynucleotide Abstracts of Molecular Radiobiology Section Publications and Manuscripts Submitted 273 1. Determination of Protein Contamination of Deoxyribonucleic Acid by t h e Folin-Lowry 2, BODPRO: An Ultra centrifugal Boundary Dlagnost i c Computer Program 3, BODPRO 11: Analysis of Tq-Bacteriophage Ultra centrifugal Boundaries 4. The Reaction of 5'-Thymidylic Acid with t h e Condensation Product of Phosphorus Pentoxide and Ethyl Ether Thymidine Derivatives on Sephadex Q 5. Effect of the Phosphorylation State of 6. Design and Construction of an Automatic Recording Mult l gradient Capillary hydropyranyl Derivatives of 3'-UrIdylic

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During this report period (FY 1984), considerable expansion of molecular and cellular level studies occurred with the arrival of new peraonnel. Projects involving clinical applications of radiation and rr dio active i.otop.8 have been brought toa reasonable conclumion, and the Clinical Invest l qr rt ions Section has been discontinued as a result of the temin8tlon of Dr. C. C. Lushbaugb. An increaming amount of the effort of the Low-Level Counting Section ham been devoted to improving electronic cell counting techniques aod developing suitable instrumentation for other sections. tions of phyaics to biological pro b lmr 8re re8pOnSible for changing the name of the section to Biophysics. These in crea a ing applicaDuring FY 1965, the categories of environmental radiation studies and health physics will be further curtailed and the effort directed toward biophyeics and in mtn u ent ation in support of the fundamental molecular and cellular level studies. During FY 1965-1968, Sfudie8 of the toxicology of radio nuclides will be continued and perhaps somewhat increased. Studies of the genetic effects of radiation will continue at a decreasing level, in view of the results obtai ned thus far, and the effort transferred to studies on somatic effects of radiation in the sub lethal dose range Construction of a new occupational health laboratory in FY 1965 for occupancy by the In du atrial Hygiene Group (8-5) and the Field Studies Group (H-8) will free considerable

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office and laboratory space Inthe Health Research Laboratory t o accommodate anticipated expansion of the biological and medical research staff. FY 1965 research activities will be devoted principally to the following program categories at approximately the percentage 06-01 Somatic Effects of Radiation (17 percent) 06-02 Radiation Genetics (18 per cent) 06-04 Molecular and Cellular Level Studies (62 percent) In addition, other projects will continue in environmental radiation studies (06-05), radiological and health physics and instrumentation (06-06), and selected beneficial app lica - The following terminations occurred during the present report P. A. Goldman, research assistant, Molecular Radiobiology N. C. Brown, technician, Mammalian Radiobiology Section. (c) Leave of Absence tion, has been granted an additional year's leave of absence t o practice medicine in the Los Alamos Medical Center. The recruiting campaign of the past two years t o fill positions created i n expanding the molecular and cellular level studies has resulted in the addition of a number of extremely well qualified personnel, New hires during FY 1964 are a s

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Carol N. Robert leculor Ra dio b lo logy lilmimm*Acceptances of Offers of Employment for work here in Sep ten wlll report for work here in

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The following table of or~pnlzrtioni nd f cates the major diVi8fOn8 of effort within the proup:

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RU'XRMIUY-106 IN MICE, RATS, AND DOGS: INTERSPECIFIC COYPARthe retention function,X is the body weight, and a and b are constant8de terri ned by le8st-8qture6 fitting, has been found t o relate t h e body weights of mice, rats, dogs, monkeys, and man tothe whole-body retention of tritium (in assumed that the well known change In metabolic r8te associated with the change in eurface-voluma relrtfonships among anilarls 02 ditiorerrt sizes is reflected in this Inter specific after oral and parent era l admin l strat lon to rats, mice, and dog8 ie awl zed here. RESULTS AND DISCUSSION Counting pmc@dures, method6 of isotope a do in lat rat fon, and treat rent of data were all routine, aa previously deFigs. 1, 2, and 3. In Figs. 1 and 2, the curves represent plots of exponential functions fit to the data by a leastand 4 dog6 are omitted for clarity. Figure 3 is a log-log plot of the integrals of the effective retention functions The results are premented in mble l and in The data points for the 12 mice, 6 rats,

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Fig. 1. Best f i t retention functions for mice, rats, and Per cent of administered dose retained I s shown a s a function of time.

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against body weight, with a least - square6 estimate of the best fit. Figures 1 and 2 show that excretion I n dogs is a slower process than in mice and rats and that mice and rats excrete ruthenium a t approximately the fame rate during the initial 400 days. The difference in route of administration (X.P. for rats and mice; 1.V. for dogs) may be a contributing factor to the difference between dogs and rodents, because excretion after intravenous and intra peritoneal dosage is not necessarily the same. The small differences among the values of the integral6for the effective rotention function (Fig. 3) suggeat that differences due t o body weight-body surface relation e do not influence the retention of ruthenium to the same degree as the retention of cesiun, iodine, and zinc are influenced by volume-surface relations, is discussed further elsewhere i n this report (6). Monkeys will be given oral and intravenous ruthenium t o increase the number of species and to afford a test of the validity of an estimate of the retention function by interpol a - Cell. Cornp. Physlol. 59, 45 (1962). W. H. Langham, Health Phys. 8, 481 (1962). Application of Regression Analysis tothe Power Function, Health Phys. (in press).

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THE RETENTION OF MANGANESE-54 BY MICE, RATS, DOGS, AND Manganese-54 was adminiutered t o mice, rats, monkeys, and dogs i n order t o determine the relation between body weights and the in te gn ls of the retention functions, formation for o t h o r isotopes has been previously reported (14) . The por sibi lit y of extrapolating from physiological parameters of mall mammals t o those of man is one of t h e major reasons for such measurements. RESULTS AND DISCUSSION and treatment of data have been previously reported (5). The results are presented in Table 1 and In Figs. 1, 2, and 3. In Table 1, the parameters of the effective retention equations are tabulated according to species and route of administration.Several elmilarlties may be seen. The I.V. rats and mice are strikingly similar, as are the oral rats and monkeys. These similarities are evident in the integrals of the retention functions (column "E. F."), which may be regarded as an estimate of the body burden in daily doses under condition e of chronic exposure or,alternatively,asa weighted half-time for all the components of the retention The slmilarltles are also ePPphaeized i n Figs. 1 and 2, where the effective retention functions given i n Table 1 are plotted. It is evident from Big. 1that rats injected intro - venously resembled mice injected intravenously more than they resembled rats injected intra peritoneal ly. That is, the difference8 due to route of injection were greater than the differences between the species. In Fig. 2, the rapid initial loss of the orally administered dose I s evident. Between 92 and 99 per cent of the dose l e lost witha half-the of less sorbed from the gut) is probably lost ina manner similar t o manganese adminiatered parent e rally. Consequently, small adjustments should be made to increase the equilibrium factors. The adjustments would be very small In the case of

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rats and mice, since less than2 petr cent of the dose is abaorbed. We have no pr ron t ~ r a ld8tr for monkeys. Only for t h e dogs would such adjurtment be porsible. In Fig. 3, the unadju8t.d equilibrium fr ctor a are plotted as functions of body weight on log-log paper, along with a least - squares reg reso i online. The line is extrapolated t o 70 kg to eat hate the body burden i n man under conditions of chronic exposure. The extrapolated value is 2.1 times the daily doae. If this is taken to equal the nmxlmum psrmissible body burden and if the total body l a taken asthe critical organ, then the (YPC)w $8 0.025 pc/ml for a 40-hour week, which does not differ from the value of 0.02 ~c given by the International Com i as ion on Radiological Protection. The agreement may be largely fortuitous in view of the fact that no adjustment of the equilibrium factors was made. It Is difficult to explain the slmilarities between rats and mice after parent era l administration of manganese-54 and between monkeys and rats after oral administration,part ic - ul a rly since this paper is predicated on finding differences related t o weight. Real differences between the excretion of m ng an ese by rats and nice are found after oral administration. The greater amount of manganese absorbed by rats passage through the rat gut (kl - 2.5) than through the mouse gut (kl - 4.9) is the principal source of difference between the handling of manganese by the two species. No such factors played a part in the parent era l administration of the isotope to rats and mice. tion rates of manganese inthe two species were insensitive tothe metabolic rate difference8that accompany abnormal environmental temperatures. In a slmilar manner, ruthenium106 was excreted at similar rates by rats and mice. Both these isotopes were refractory tOW8rdthe enhancement of excretion(6)with the ambient temperatures shown by cesium137 (7) and strontium-89 (8). It may be postulated that less "labileft elements will show no differences In excretion rates between closely related species such as rats and mice and Will be less susceptible to change by metabolic stimuli or The factors controlling the excre-