Quarterly Technical Report July 1, 1952 Thru September 30, 1952

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Health and Biology Atomic Energy Project Rochester 20, Nev York July 1, 1952 thru September 30, 1952 It should be noted t h a t the Quarterly Technical Reports of The University of Rochester Atomic Energy Project do not attempt t o describe progress inall of the research programs but only i n those i n which some significant results have been achieved but which are not sufficiently comilete t o be mitten up as Submitted by: Henry A . Blair Date of Report: 11/3/52

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TABTZ OF CONTENTS Rabbit as Measured by Tolerance t o Pyrogenic Material 4by R.W. Miller and R . Watson ................. EXTERNAL RADIATION - TEIERAPY The Effect of Whole Blood Transfusion on Dogs Receiving an Do Exposure t o Ionizing Radiation by W. K . Cotton "he Effect of Crcss-Transfusions on the Acute Rsdiation Syndrome i n the Dog by John L. Grace, Joseph Seeger and Studies Relative t o the Effect of Piromen on the Number and t h e Phagocytic Activity of Leukocytes of Irradiated and Normal Dogs, and of One Man by M. Ingram, M. Adams, A New and Simple Differential Morphology Stain for Spermatozoa by George Id. Casarett ............... 37 Studies of Flash Burns: The Influence of Spectral Qualities of the Carbon Arc Source by Kelly M. Berkley, Herman E . ABS!FIACTS OF PAPERS PRESSNTED AT MEETINGS Application of Some Multivariate Anal.ysis Techniques t o Data fron Radiation Experiments by Arthur M. Dutton. ........ 50 Blood Fluoride Concentrations i n Dogs Exposed t o Hydrogen Poisoning by Charles J. Spiegl. ................ 52 Selectzd Blood and Urine Changes i n Experimental Beryllium

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Urinary Excretion of Sodium Fluoride Administered in t h e Cyclctron Zlazzrds and Thc3.r Managemcnt by Herbert Mernzgen . . 56

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9 rabbite were mde to le rapt to the daily intravenous adminiatratlon An LDW of tota lof typhoid Taccine as measured by their febrile response. bo@ irradiation does not abolish this tolerance. reticuloendothelial system to this tolerance is briefly discwsad. The rel a tio whip of the Rabbit8 develop a tolerance t o pyrogens as msasured by body temperature. It has been suggested by Beeson (1)t hat this tolerance it3 a function of the intravenous typhoid vaccine is abolished by a Thorotrast blockade of t h e rsticuloendothelial system. If Beeson's thesis is correct, then a f'unctional disturbance of t h e r-c system produced by x-irradiation might be demonstrated by a loss of body temperature tolerance t o typhoid vaccine. Mathods and m te rials : He bases t h i s concept on the fact that tolerance to A.6 healthy male albino stock rabbits were wed. individuel cages i n e. room undar constant temperature and hwnidlty. They were kept in The animals were placed i n wooden boxes for the periods when their tempcrar;uras were being recorded. circuler piece reu?oved atits anterior end t o hold the r sbb it ' s head outside The sliding top of the box had a semiThe r e a r pane1 of the box flapped open on hinges, thus allowing t h e 1 -? + -rectal temperature t o be obtained with a minim disturbance to the rabbit.

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Ppogen b e gla-eftware and needlea were employed. These were checkad from time to time by using them to inject pyrogen-free saline intravenously i n U. S . Public Health typhoid vaccine containing one billion organisms per ml was used as the pyrogenic material. The same lot was wed throughout ~ lii swas kept under continual refrigeration and shaken vigorously bafore'ase. It was diluted 1:lO with physiologic saline (100 million The enimls were placed .in the wooden boxes for 4-6 hours tllzi4 for the two days prior to the first indoction of typhoid vaccine. Thfa was done t o accustolri them t o the procedure. Baseline rectal temperature fluctuations were recorded during this time. Each day after a baseline temperature had been recorded, each rabbit wag 'given one cubic centimeter of typhoid vaccine into the marginal vein of She ear. Its rectal temperature w m then recordad every half' hour, u n t i l it -returned t o normal. The thermomters were inserted in the rectum for one ninute by the clock for each recording. The same them meter was used for a11 rabbits at all times. 800 r 250 Kv. Lead plano-convex filter, 1/2 inch at center, 1/4 inch a t periphery. I@: 15, TSD: 44 inches, Rate: 6.9 r.

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B . 3 healthy adult albino rabbits were w e d under the earn experimental circumstances, but t h e treatment schedule was altered: Dag of experiment 1 2 3 4 5 6 7 8 9 i o 11 12 13 14 15 16 17 * Total body irradiation after temperatures were recorded. * Total body irradiation after temperatures were recorded. Part A . The rabbits davelopcd a tolerance t o pyrogen which was not altered by a n L;D50 of total body x-irradiation when the pyrogenic material wa8 given daily except f o r the day af'ter t h e x-ray exposure (Graph I). The rabbits developed a tolerance to pyrogen which was not altered by even though a brief lapee i n the pyrogenic stixulw was allowed. Thus, the tolerance was maintained Bloom ( 2 ) describes large amount6 of lymphocytic debris i n t h e phagocytes of rabbits 17 hours af'ter total body x-irradiation. This euggeets that the reticuloendothelial system may have been blockaded by the debris. et. al. showed that 16-18 hours afier an intravenous injection of this drug no radioactive gold could be phagocytized. Following x-irradiation, on the other hand, the sama method of testing showed no impairmbnt of ability to phagocytize. This indicates that this function of the r-e syetem 18 not affected by x-

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Beeson showed that int.ravenous Thorotrast injection in rabbits abolishea t h e i r tclerance t80 pyrogem (11, However, Thorotmat disturbs the physiology top pog cns is the consequence of r-e blockade i s based as yet on inconclusive t h i s actlon when Thorotrast is administered, it mst be asswned that a similar Therefore, seeson's claim that. the abolition of tolerance Hovever, if a reticuloendothelial blockade is solely respomible for blocbade does not occur when amb bit is exposed t o an T , ~ Oo f total body xits abolition by reticuloendothelial blockade, Proc . SOC. =per. Biol. Bloom, W.., Histopathology of irradiation from external and internal reticuloendnthelial syote ~ 2(RES) i n rzi3bits Injected intravenously with

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Ten o f the63 aiim19 served as controls, while the other t e n received typed, cross-mtc.ned. comp8.tible whole blood tranefbsions on a predetermined schedule during t h e post.-imzdiation test period. There were no deat;”; in either group. It would seem, therefore, that the f’regueEt administraticn o f therapeutic amounts of‘ whole blood i s not detrimentel t o tha survivel ofa dog thzt has received a non-lethal exposure to ion iz iw irradiation. This exyeriment w2s bne as consequence of a study by J. G. Allen, e t a1 ., whose results suggested a substantial increase i n mortality resulting from t h e R-oquent adzirniskratim of whole blood t r J dogs receiving an 1% exposure t G icnizicg rrradiztior,. InsroaucTcic3r: J . Gammt Allen a l ~h i s aseociatss have published a sa.% i;? b o t h cno control and the t.ransfused groups.

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I 3 seemed appropriate t o repeat a determine If the yspeated administration portion of this study i n order t o cjf therapeutic quantities of whole blood is a primzry hazard toa subject Methods : Tienty unanesthetized as possible i n respect t o weight, sex, mongrel dogs were pzired as closely age, and blood type. Each animal was celved typeu, cross -matched, compatible whole blood on a predetermined basla of' thee transfusions a week beginning on the4th day poet-irradiation for 48-72 hours prior to its administration and stored at hoc. into the leg vein inths amount of5 cc/kg. of body weight. wa8 withdrawn from each animal 3 xiweekly i n order t o follow its he m to crit, REC, WBC and platelet counts. i'ollowing irradiation served as a control while the other 1'8The blocd waa drawn from donor ani mls in standard ACD sOlUtiOn A fourteen dog donor colony was maintained. Each donor and recipient animal vas typed for the presence o r absence of the do6 "A" factor. was CrOES-mii.t,che&for mjor and mincr incompatibility just prior to transfusion. The diet 02 the & mor zoloqy was supplemented with hily feedings of mat and irm.which mint a in ed the donor hematocrits a t approximatelyhorse 95% of their contrcl lave is. 1. hl or tal it y - None o f *he dogs succumbed during the 28 day experi2 . Hemtolorn - In both the control and transfused g o u p s , the leucocytes and platelets fell off rapidly during t h e first ten days af-cer irradiation, reaching a minimum level

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by the fifteenth post-irradiation day. minimum f o r both groups, as shown i n Graph I, was the platelet count ina few bga reached zero, and afew control doe had a WBC count o f 400 fora two-day period. Graph 111 shows that the red cell count ofthe tranafuaed group remained atthe pre-radiation control level while of both groups fell during the initial three day period until t h e first transfuaton date when the hematocrit of ths transfused group rose steadily to a level higher . than its ncrmal control value. This l e v e l waa then maintai ned until the end of the test period. The hematocrits of the control group paralleled t h e RBC's by falling gradually and cornlatently throughout the four week period. 3 . Blood trans hio m - Bo clinical hemolytic o r anaphylactoid transfusion reactions were noted. t o corroborate the findin@ of J. G . Allen, e t al., as given inthe introblood given toabg which has been exposed toa non-lethal dose of xirradiation is detrimsntal tohis survival. our results indicate no effect on mortality. The parallel fallin WBC's and platelets indicates a primary bone marrow defect as a r e s u l t of irradiation which was not overcome by transfusing with whole blood. The results of our experiment under the stated conditions fail Since all of our animals survived, w e cannot state that whole In attempting t o explain the varying results obtained from two baaically

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similar studies done by Allen and ourselves, an interpretstion l e offered i n the differences existing between the dosage of radiation and the minimum leucocyte count. Allen's 175 r appears t o be the highest dose of total body x - irradiation that can be given toa group of dogs inhis laboratory without producing death. A corresponding effect In o w laboratory, usiw different radiation factors, is produced by 300 r. by the leucopenia resulting from the Irradiation in the two experimenta, but Allt3n'e data for this is not available in the literature. The effects might be compared further

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THE EZTECT OF CROSS-TRANSFLTSIOI’lS ON TEE ACUTE John L o Grace, Joseph Seeger, and Robert w. mller 12 dogs were given an r;~8oof t o t a l body x-irradiation. 6 of these animals served as controls. post-irradiation day, and 3 were so treated from the 7-11th post-imadiatioa norm1 animal, the same donor being wed thro-out the five day period. procedure did not diminish t h e mortality. hematology of t h e peripheral blood was detected. Each cross-tran~fueeddog received its tranefueion froma compatible Eo consistent major change inthe The work of Salisbury, Rekere, e t a1 (1) suggests t h a t croas transfusion lowers the lethal it y of x-irradiation. Swisher and Wh(2)per fo md a small exchange transfusion (250cc) using f’resh whole blood on dogs immadiately af’ter they had been irradiated. Their results fiom a slnall number of animals suggestod t h a t the mortality was diminished. O n the other hand, stored blood had no effect on the lethal it y of total body x-ray exposure (3). Because these studies showedthat fiesh blood m y be beneficial i n radiation siclmess and becawe of the hopes provided by the results of the cro ~ s transf’usiorm mmtioned above, an extension ofthi8latter work waa carried out. the same uel gh ts were studied concurrently as follows: 18 healthy adult bo@ of mixed breed were w e d . 3 dogs of approximately

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6 groups o f dogs were studied in t h i s way. 5OOcc of blood were crosatransfksad every day for five days. received their transf'usiom from the l-5th day posi-irradiation while the other Three of the irradiated treated animal8 These shimla hcd been immunized for distemper and given a vermifuge approximately six weeks befora we. They were kept i n separate cages and fed Ediet of Wina dog chow with free access t o water both before and during Control- &tarmin&tions of hematocrit, red blood cell count, white blood cell count, pltltelet couit and blood smear were lnada on each of3bys of the week pri cr to irradiation. Folloving irradiation, these blood studies were made on week (3ap for the first 2 1 days und three times foy the next week until the 28th day. Peri-Pheral blood studies word also mabs on som, but not all of the dogs a f t e r each cross tmnaf'usion. standard laboratory methods were uaed for all these &eterminations, The animls t o be cross transfkssd were typed for presence of the canine "A' factor in t h o i r erythmcytes a8 described by Christian, e t al. ( 4 ) and Yew, et al. ( 5 ; G c r q ~ t - i b kdonors and recipients were chosen 80 a8 t o Donor ' 6 cslls and recipient '9 serum, and recipient s c e l l s and donor's Berum -- venipunctaree were carried out using #13 needle inaerted in either rip ht or lef% ,jugular vein end e #l9O polyethylene catheter placed into the vein for e distance fYom 1.0 t n 15 cm. (depending on the size of the dog),

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The cathetsrs from thot wc dogs were attached toa three-way stopcock by maria o f Luer-Lok Couplers and a 20cc syringe was connected tothe stopcock for exchanging the blood i n 20cc quantities until lOOcc had been croes transfueed i n each direction. All aDparatus was lined with silicone previous t o each cross The pairs of animals t o be radiated, one of which served a6 t h e r e - cipient and t h e other as a control, were exposed t o 3wr (approximately I;D80) delivered froma 1000 m General Electric Industrial X-ray unit operated at 3 ma wing a lead plano-convex filter6 inch thick in the center. Radiation was delivered a t a rate of approximately gr/minute as measured by a Pictoreen …