The Effect of Graded Doses of Ionizing Radiation on the Human Testis

Page 1

IST?.ODUCTION AND PROJECT HISTORY At a meeting held ir.Fort Collins, Colorsbo, in1963,a discussion arose about objective knowledge of the effects of radiation in non - lethal doses upon the human body.Since as to hcn to obtain it. was the only logical internal organ that could be irradiated exterior to the rest of the body, without surgery land that would. be sensitive to sra all changes.Ina ~ d it ion ,t he testes would offer direct information on reproductive organ effects, a prime area of interest. I t was decided that the male yonad Subsequently,a proposal was submitted to initiate such a study.The proposal was approve2 and begun in the late summer of 1963. .Advisory con witte e nec tings attended by various scientists and AEC personnel were held in1963,1965 and 1967 to critically review all facets of the project and volunteer handling and to offer advice as to future procedures. Soze members of the committees visited the penitentiary, viewed the laboratory arrangements there,and talked with the

Page 2

PRIVACY ACT MATERIAL REMOVED X-ray irradiation of the volunteers was performed using a unit constructed irradiated a second time and one subject a third time. irradiation procedures were performed following complete recovery. received 5 rad/week for 11 weeks,for a total of 55 rads. Irradiation s were begun on t22, 1963 and finished on &ring this time, 67 men were irradiated. O f these, six were The second and third Completion of the follow-up on those irradiated during later years was interrupted early i n 1973 due t o the total medical incapacitation of the then principal investigator, and the non-acceptance o f the State of Oregon Director of Correctional Institutions to allow another medical doctor t o continue as. supervisor of the program (due to fear of possible increased publicity, and pol it ical and legal manifestations). Minimal follow-up was allowed by those of us already involved w i t b the , volunteers -and long-term medical supervision is s t i l l being provided by Dr. Daniel Dilaconi, thersurgeon who performed allof the testicular.biopsies, capacity, the Principal Investigator was changed on February 2 2 , 1973) and evaluation o f all - data .has<been re-coded, s o no reference can .be found which will i n any :.

Page 3

manner identify any o f the volunteers. cular biopsy slides, tissue blocks and samples collected for electron microscopy. Sperm samples and plasma samples have not yet been re-coded. After the closing of the laboratory a t the Pacific Northwest Research Foundation,. all specimens and data were relocated i n the Division of Nuclear Medicine inthe Tumor Institute of The Swedish Hospital Medical Center. The transfer (from Pacific Northwest Research Foundation toa storage unit to Swedish Hokpital; or from Pacific Northwest Research Foundation t o Swedish, mati on, and other information o f potentially legal significance have been lodged i n two safety deposit boxes i n the First Hill Branch of the Rainier A1 1 consent forms, psychiatric evaluation infor -

Page 4

sclcction of Subjects The 67 subjects for- this study were healthy noc nal men between the ages of 25 and 52 years. Their reproductive normalcy was determined prior to irradiation by physical exam in a Gion,examination of testicular biopsy specimens, sperm concentration and sperm morphology. and testosterone were also determined.Additional routine tests S: a s blood cytology,urinalysis and various blood chemistry tests were evaluated during the pre- and post-irradiation period. Levels of urinary total A Device to Deliver Uniform X-Ray Doses deter xi ned dose of X - radiation throughout both testes of hunan independent of plumbing and with built - in radiological protection. EXF O SUre doses, uniform to within 2, 5E.of the stated average were t o be delivered at the rate of approximately 100roentgens operating at approximately 140 KVP,with up to a 5 m-a. tube

Page 5

current,having an inherent filtration of 0.14" glass plus These units were mounted in a lead - lined box with their beams horizontally opposed ona corrunon axis. The X-ray beams were incident on a block of tissue equivalent material20 crns 1n sert ed.into this block of material in the middle of the upper 94'F ) water which served two purposes, obtain o ' esc cnt of the testes into the vessel,and making the testes,water vessel and surrounding material one tissue equivalent mass,there by making the dosed is tri but iGn in the testes independent of the natural variations in the In e ppl,i cz tion,the cup was filled with water at s cro tel temperature and the subject laid prone with testicles suspended in the water. The level of the teskcl'es i n the cup was viewed in the mirrors through the view ports,.. When the testicles were beam path and the pre de term in ' ed dose delivered. The unit was re calibrated at the termina tic n of the ex per irn ent. show less than 1%c han gein output during the run.

Page 7

One independent check of the X-ray calibrations was made . 1 using Rlrneters by a physicist at Bat tell e Northwest Laboratories. i:i He don firmed.that " Our independent calibrations agreed with those of the experimenters within the accuracy expected from the results was about 32. " A second independent check was na de using a biological standard, by Dr.Eugene F. Oakbcrg of Oak Ridge h ' at ional Laboratory. would fit into the water box of the X-ray machine was ,constructed.. Dr.Oak berg ' s genetically pure strain of--nice was used. were irradiated at various dosages, then autopsied at :72 hours - A special mouse - holding cliainber that after X-ray. The preserved testicular spec hens were analyzed and the results compared to results obtained using equipment a t1

Page 9

Data on Ir x Adi ation of Subjects Sisty - seven subjects received acute testicular ir radi a tic n in doses between 8r and 6OOr. Six subjects were irradiated a secon8 time, ' and one subject a third time,following complete recovery from the initial dosage. O f these, three received the identical dosage on each of the two occasions,three received two different dosages, and one received three different dosages. One subject was qiven weekly irradiation s of S r ?er week for ' eleven weeks. Each dosage given is listed separately i n T'A!3LE 1 1 For each paraineter studied each subject served as his Post - irradiation observations were Gnde on kach mentioned above. Serial test icu lz r biopsies were during the post - irradiation period i;r orcier t o specifically evaluate sperm snd hormonal evaluations 1-5 tabulated as one irradiation only..).All 'biopsies were prepared I for light .microscopy and inthe.3ater.years of the - study I for electron microscopy. Germinal cell and Leydig cell quant it ation methods were developed and utilized on. fixed for light microscopy.._

Page 10

Sperm counts were performed using the ha emo cyto meter iRcthod on samples submitted weekly by each volunteer. During the first years or' this study,urinary.st c roid and hormone evaluations were used ex clu sib ely. became available,urinary evaluations were phased out. urinary gonadotropins were concentrated using the speed - filter and kaolin adsorption method of Albert.Totai gonadotropins were bio assayed by the method of Laus on, Heller and S ev ring haus.Luteinizing hormone bioassay was conducted using the male hy pop hy sect on ize dr at method of Grecp, etal. -.. The Steel man - Pohl ey essay '.- was c sed for As plasma methods Estrogen s were bio - assayed using the net hod of Laus on, and epi test ost crone levels were determined using gas - liquid Plasma testosterone was deter yi ned using the method of and protections.were designed and implemented throughout this entire program is attached as APPENDIX B.'

Page 11

A brief overview of the findings Pcblished i n Radiation Research entitled " Effect of Graded

Page 14

Superscript numbers l.through6 refer. to the six subjects who received...- more than one X-ray dose, 'e.g.. both 25r doses were received by the tame

Page 15

geWZfS: an ert i mate of the duration of each cell association and of each cell type. 111 International Ccngress : Of Endocrinology, J u n e 30 - July 5 , 1 9 6 0 , Mexico City, Excerpt Iledica International Congress Series,184, 1012-1018, 1563. of sperrnat~~zoathrough .the human male duct u lar system. sem in if ero us epithelium of the human testis cfilploying the Sertoli cell as a constant. 2. Zellforscll. , lis, 461-472, Exccrpta Nedica Foundation, International Congress Series, Erusca J Action of radiation upon human rpermstogcnesis. . I method for the quant if i cation of Leydiy cells in man. 3. concentration due to testicular biopsy procedure i n man. < weights of mature ova rie c to miz ed rats. Cndocrinology, 21, graded .doses of estrin upon the pi tui tar),,adrenal and thy nus