List of Experiments

Hanford Sites

HS-1. Ingestion of Iodine-131 in Milk by Hanford Employees

IN 1963, milk from dairy cows fed iodine­131 (I¹³¹) was consumed by eight General Electric/Hanford workers either as a single dose or as several daily doses. During the study, the amount of iodine in the cows=diet was increased from 5 milligrams per day to 2 grams per day. The resulting uptake by the human thyroid was determined in Hanford=s whole-body counter facility. Participants were Hanford scientists who had volunteered to drink the milk and be counted over a period of approximately 1 month. This work was supported by the U.S. Atomic Energy Commission. (Previously described in #41 on the original list of 48 experiments released by DOE in June 1994 and included in The DOE Roadmap of February 1995)

References

Watson, E.C., I.C. Nelson, D.H. Wood, R.O. McClellan, and L.K. Bustad. AEffect of Varying Stable Iodine in Diets of Cows Fed I¹³¹ on Uptake of I¹³¹ in Man Drinking the MilkCAn Abstract.@In Biology of Radioiodine: Proceedings of the Hanford Symposium on the Biology of Radioiodine, Richland, Washington, July 17B19, 1964. Oxford: Pergamon Press, 1964, p. 339.

Handwritten Monthly Report. J.K. Soldat to R.F. Foster. July 1963. Pasco, WA: Washington State University Tri­Cities Campus, PNL, DOE Richland Public Reading Room, I¹³¹, Open Shelving, PNL­936 9­DEL.  "

HS-2. Intentional Release of Iodine-131 at Hanford in 1963

IN JULY 1963, the Hanford Laboratory conducted a study that involved the release of 120 microcuries of iodine­131 (I¹³¹) into the environment. These releases were designed to characterize the dispersion of radiation. The purpose of the experiment was to enable scientists to determine the fraction inhaled by men, the amount taken up by the thyroid, and the retention half-time of radioiodine in human thyroid.

Two volunteer subjects (Hanford employees), were stationed in the expected path of the radiation cloud. These subjects intentionally inhaled I¹³¹ from the release and were subsequently measured for thyroidal uptake of I¹³¹. These experiments were performed under contract with the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995, and since revised)

References

Gamertsfelder, C.C. APlans and Hazard Analysis for the First Hanford I¹³¹ Field Release Test.@Richland, WA: Hanford Atomic Products Operation, Physics and Instruments Laboratory, HW­78312, July 19, 1963. Pasco, WA: Washington State University Tri­Cities Campus, PNL, DOE Richland Public Reading Room, I¹³¹, Open Shelving.

Handwritten Monthly Report. J.K. Soldat to R.F. Foster. July 1963. Pasco, WA: Washington State University Tri­Cities Campus, PNL, DOE Richland Public Reading Room, I¹³¹, Open Shelving, PNL­9369­DEL.

Monthly Report. Senior Engineer to R.F. Foster. August 23, 1963. Pasco, WA: Washington State University Tri­Cities Campus, PNL, DOE Richland Public Reading Room, I¹³¹, Open Shelving, PNL­9370.  "

 ‡  ‡  ‡

HS-3. Absorption of Tritium Oxide Through the Skin

IN 1951, scientists in the Biology Section, Radiological Sciences Department, General Electric Hanford Company examined the percutaneous (through the skin) absorption of tritium (H³) oxide-labeled water vapor in animals and humans. A total of 14 subjects participated in the study.

Twelve subjects were exposed over about 10 square centimeters on the forearm, and two were exposed on the abdomen. One of the arm-exposed subjects also received atmospheric whole-body exposure while breathing H³-labeled air through a respirator. Urine samples were collected from all 14 subjects over a 48-hour period.

The study results indicated that absorption of H³ through the skin and lungs from an H³-contaminated atmosphere contributed similarly to the total-body burden as exposure by H³-water vapor. As a result of this finding, the researchers recommended a 50 percent reduction in the permissible maximum level for atmospheric tritium oxide. This study was funded by the U.S. Atomic Energy Commission. (This experiment was referenced in the Markey report.)

References

DeLong, C.W., R.C. Thompson, and H.A. Kornberg. APercutaneous Absorption of Tritium Oxide.@American Journal of Roentgenology, Radium Therapy and Nuclear Medicine. Vol. 71, No. 6, June 1954, pp. 1,038B1,045.  "

HS-4. Calibration Studies Using Zinc-65

IN 1959, researchers at the General Electric Hanford Company, Atomic Products Operation conducted studies on the retention and distribution of zinc-65 (Zn⁶⁵) in the human body to establish calibration values for bioassay and in vivo counting.

In September 1959, one subject (a Hanford worker) was orally administered an unstated amount of Zn⁶⁵ and the amount remaining in the body was measured by whole-body counting. The uptake of ingested Zn⁶⁵ was found to be 35 percent or about twice the amount expected from results of animal studies. Over time, different values for different individuals suggested that different distribution patterns should be considered.

In March 1960, a study involving one subject showed that Zn⁶⁵ deposited initially in the liver and later in the head and legs. The General Electric Hanford Company was operated at Richland, Washington, for the U.S. Atomic Energy Commission.

References

Bioassay Laboratory Monthly Report, SeptemberB1959.Richland, WA: General Electric Hanford Atomic Products Operation. October 1, 1959.

Radiation Protection Operation Report for the Month of March, 1960. Richland, WA: General Electric Hanford Atomic Products Operation. April 11, 1960.  "

HS-5. Study of Metabolism of Strontium Using Strontium-85 as a Tracer

A STUDY WAS CONDUCTED in 1963 by scientists of the General Electric Hanford Laboratories to (1) determine whether ingested strontium is excreted in human body hair in measurable amounts, (2) determine whether analysis of hair samples was an accurate indicator of strontium uptake in man, and (3) investigate the biological retention of ingested strontium in man.

Two Hanford scientists voluntarily ingested a solution containing a few microcuries of strontium-85 (Sr⁸⁵). The exact amounts administered are not known. Hair clippings and facial shavings were then obtained from the subjects and analyzed for Sr⁸⁵ content.

The results of this study showed that Sr⁸⁵ could not be measured in small samples of body hair. This study was discontinued and the results were never published. The Hanford Laboratories were operated by the General Electric Hanford Company for the U.S. Atomic Energy Commission.

References

Letter. T. Beasley to D.R. Fisher. May 29, 1995.

Letter. B.I. Griffin to D.R. Fisher. May 2, 1995.

Memorandum. D.R. Fisher to file. April 27, 1995.   "

HS-6. Whole-Body Counting Studies of the Measurement of Phosphorus-32

DURING 1963 TO1964, a researcher at the General Electric Hanford Company developed techniques for the whole-body measurement of phosphorus-32 (P³²). The purpose of this study was to develop a more sensitive method for measuring the level of beta-emitting radioisotopes in the body.

Three patients at the University of Oregon Medical school who had received P³² therapy for blood diseases participated as subjects. Two other volunteer subjects at the Seattle Swedish Hospital were administered P³² by injection for calibration study purposes only. Each of the subjects was counted for P³² in the Hanford whole-body counter.

These studies provided P³² calibrations for measurement equipment that later was used in studies of P³² in people who consumed fish from the Columbia River. This work was supported by the U.S. Atomic Energy Commission.

References

Palmer, H.E. ADetermination of P³² In Vivo.@Health Physics. Vol. 12, 1966, pp. 605B608.  "

HS-7. Distribution and Excretion of Technetium-95 and Technetium-96 in Humans

IN APPROXIMATELY 1965, scientists at the Battelle Memorial Institute=s Pacific Northwest Laboratory collected data to establish exposure limits for personnel engaged in the handling of technetium isotopes and for workers in areas where inadvertent exposure might occur.

Eight normal volunteers, between the ages of 23 and 40 years, received 20 microcuries of technetium-95 (Tc⁹⁵) and 60 microcuries of Tc⁹⁶ as pertechnetate. Administrations were by intravenous injection to four subjects, and orally to the other four. Samples of blood, urine, sweat, tears, and intestinal mucosae (biopsies of stomach, duodenal, and rectal mucosae were obtained from three individuals) were collected for the first week; total urine and feces were collected for an additional 8 to 10 days.

The study results indicated that there was no detectable difference between the intravenous and oral administrations in relation to the rate and route of excretion, and the amount excreted. Technetium was found to concentrate primarily in the stomach and gastrointestinal tract, with little or no concentration in the liver or kidney. Blood and urine analyses together were found to provide the best method for assessing external depositions of technetium as pertechnetate following accidental exposure.

The researchers concluded that the recommendations of the International Commission on Radiological Protection on the maximum permissible body burdens and air and water concentrations of technetium as pertechnetate, needed to be reevaluated. This study was funded by the U.S. Atomic Energy Commission. (This experiment was referenced in the Markey report.)

References

Beasley, T.M., H.E. Palmer, and W.B. Nelp. ADistribution and Excretion of Technetium in Humans.@Health Physics. Vol. 12, 1966, pp. 1,425B1,435.  "

HS-8. Study of Promethium Metabolism and the Effects of DTPA on Promethium Metabolism

IN 1967, SCIENTISTS at Battelle Memorial Institute=s Pacific Northwest Laboratory and the Hanford Environmental Health Foundation, conducted a collaborative study on the effects of DTPA (diethylenetriaminepentaacetic acid, a chelating agent) on the retention of promethium-143 (Pm¹⁴³). The purpose of the study was to develop an excretion model for diagnosis of promethium exposure, to form a basis for radiation exposure, and to determine the radiation dose from accidental exposures.

A total of 14 volunteers participated in the study. Six received approximately 0.1 microcurie of Pm¹⁴³ intravenously; two received 10 microcuries of Pm¹⁴³ orally; the final six received 0.1 microcurie of Pm¹⁴³ intravenously, followed at various intervals by intravenous administration of 1 gram DTPA to study its effectiveness in combining with promethium to enhance its rate of excretion. Whole-body counts and excreta measurements were conducted for up to 1 year following these administrations.

Results indicated that DTPA is effective in significantly reducing the body content of promethium when administered within about 1 hour following its entry into the bloodstream. This study was funded by the U.S. Atomic Energy Commission. (This experiment was referenced in the Markey report.)

References

Palmer. H.E., I.C. Nelson, and G.H. Crook. AThe Uptake, Distribution, and Excretion of Promethium in Humans and the Effect of DTPA on those Parameters.@Health Physics. Vol. 18, 1970, pp. 53B61.  "

HS-9. Whole-Body Counting Studies Using Iron-59

IN THE LATE 1960s, researchers in the Department of Medicine, University of Washington, Seattle and Battelle Memorial Institute=s Pacific Northwest Laboratory conducted studies to demonstrate the utility of the whole-body counter for measuring iron-59 (Fe⁵⁹) in humans and to establish the quantitative validity of measuring Fe⁵⁹ absorption and loss by this technique.

Several healthy subjects were orally administered 1 microcurie of Fe⁵⁹. Whole-body counts were obtained immediately after the administration and at 3 to 7 hours thereafter. Subjects then received 1 microcurie of Fe⁵⁹ by direct intravenous injection. Whole-body counts were repeated and compared for consistency. Negligible differences were found between the measurements of interests obtained following oral administration and those obtained following intravenous injection.

Another study was conducted to assess the differences in counting efficiency for Fe⁵⁹ circulating in the blood, and the Fe⁵⁹ localized in the red bone marrow. The researchers were supported by the U.S. Public Health Service and the U.S. Atomic Energy Commission.

References

Palmer, H.E., J.D. Cook, K.G. Pailthorp, and C.A. Finch. AA Whole-Body Counter for Precision In Vivo Measurement of Radio-Iron.@Physics in Medicine and Biology. Vol. 15, No. 3, 1970, pp. 457B465.

Palmer, H.E., J.D. Cook, K.G. Pailthorp, and C.A. Finch. The Precision In Vivo Measurement of Radioiron by Whole-Body Counting. Seattle, WA: University of Washington, Department of Medicine.  "

HS-10. Alternative Method for Measuring Iron Uptake Using Iron-59

A STUDY WAS CONDUCTED in 1969 at the Department of Medicine, University of Washington, Seattle in collaboration with Battelle Memorial Institute=s Pacific Northwest Laboratory on three normal subjects between the ages of 25 and 40 years.

One microcurie of iron-59 (Fe⁵⁹) as ferrous sulphate in water was administered orally to each of the subjects. Whole-body counts were obtained immediately following the oral ingestion, and again 3 and 7 hours later. The latter count was immediately followed by an injection of Fe⁵⁹ in the form of sodium citrate bound with fresh plasma. The experiment was conducted to evaluate a new (about 1970) technique for whole-body counting, namely the use of longitudinal scan geometry. This technique was tested using different biological distributions of Fe⁵⁹ in the body, including point sources, moving sources, and metabolical sources.

The study demonstrated that the longitudinal scan geometry was suitable for measuring radioactivity in humans. The research was supported by grants from the U.S. Public Health Service and the U.S. Atomic Energy Commission.

References

Cook, J.D., H.E. Palmer, K.G. Pailthorp, and C.A. Finch. AThe Measuring of Iron Absorption by Whole-Body Counting.@Physics in Medicine and Biology. Vol. 15, No. 3, 1970, pp. 467B473.

Letter. C.A. Finch to S. Marks. July 17, 1984. Pacific Northwest Laboratory General Human Subjects, Box Alan Rither, PNL-9055-DEL.  "

Idaho Sites

IS-1 Administration of Radioactive Material to Volunteers to Test or Calibrate Analytical Equipment

FROM 1965 TO 1972, as many as 18 employees at the U.S. Atomic Energy Commission=s Health Services Laboratory at the National Reactor Testing Station in Idaho voluntarily swallowed radioactive material or inhaled radioactive noble gases, prior to being placed in whole-body counters. The following radionuclides were used in the experiments: argon-41, potassium­42, manganese­54, cobalt­60, zinc-65, krypton-85m, zirconium-95/niobium-95, ruthenium-106, silver-110m, iodine­131, cesium­132, xenon-133, cesium­137, and cerium-144.

In most of the ingestion cases, the radioactive material was encapsulated in plastic so that no radioactive material was absorbed into body tissues. These measurements were performed to develop and evaluate new whole-body counting equipment and to calibrate that equipment. The whole-body counting equipment was used to measure the amount of radioactivity inside the body of occupational radiation workers exposed to radioactive material. Policies for conducting these experiments limited radiation doses to volunteers to levels below the occupational radiation­protection guidelines in effect at the time. (This experiment was referenced in the Markey report and included in The DOE Roadmap of February 1995.)

References

Anderson, J.I., and D.G. Olson. AA Rotational Technique for Assessing Quantity and Distribution of Body Radioactivity.@Health Physics. Vol. 13, 1967, p. 719.

Anderson, J.I., and D.G. Olson. AComputerized Helical Scanning to Determine the Location of Specific Nuclides in the Human Body.@Health Physics. Vol. 23, 1972, p. 325.

Howard, L.E., J.H. Spikard, and M. Wilhelmsen. AA Human Radioactivity Counter and Medical Van.@Health Physics. Vol. 21, 1971, p. 417.

Olson, D.G. AA Direct Calibration Using Gamma Spectrometry for Measuring Radioactivity in Humans.@Health Physics. Vol. 14, 1968, p. 438.

Sill, C.W. Some Guidelines for Studies Involving Internal Administration of Radioactive Materials to Human Volunteers. Idaho Falls, ID: Idaho Operations Office, U.S. Atomic Energy Commission, IDO­12058, October 1966.  "

IS-2. Controlled Environmental Radioiodine Tests (CERT)

U.S. ATOMIC ENERGY COMMISSION scientists and other professionals at the National Reactor Testing Station in Idaho conducted the Controlled Environmental Radioiodine Tests (CERT) to study the transport of radioiodine through the air­vegetation­cow­milk­human food chain from 1963 through 1968. Five of the 24 CERT tests involved exposure of volunteers to iodine­131 (I¹³¹) to study the transport of radioiodine to and through the human body.

In the first test, CERT No. 1, seven individuals consumed milk from a cow that had grazed in a pasture where the radioiodine was deposited, and their uptake of radioiodine was determined by thyroid gland monitoring. The average thyroid dose was 0.39 rad; the maximum thyroid dose was 0.63 rad.

In CERT Nos. 2, 7, and 10, three individuals, seven individuals, and one individual, respectively, were reportedly exposed during radioiodine releases over the pasture to determine their intake of I¹³¹ by inhalation.

The number of individuals involved in a similar inhalation experiment during CERT No. 11 was not listed in published reports; however, whole-body counting logs indicate that 10 individuals were apparently involved.

Thyroid doses from inhalation during CERT No. 2 were no greater than 0.015 rad, and the reported thyroid activity observed during CERT No. 7 was about the same as that in CERT No. 2. Thyroid doses to volunteers were not reported for CERT Nos. 10 and 11. The volunteers were employees of the U.S. Atomic Energy Commission. (This experiment was referenced in the Markey report and included in The DOE Roadmap of February 1995.)

References

Bunch, D.F., Editor. Controlled Environmental Radioiodine Tests Progress Report Number Two. Idaho Falls, ID: Idaho Operations Office, U.S. Atomic Energy Commission, IDO­12053, August 1966.

Bunch, D.F., Editor. Controlled Environmental Radioiodine Tests, Progress Report Number Three. Idaho Falls, ID: Idaho Operations Office, U.S. Atomic Energy Commission, IDO­12063, January 1968.

Hawley, Jr. C.A., C.W. Sill, G.L. Voelz, and N.F. Islitzer. Controlled Environmental Radioiodine Tests at the National Reactor Testing Station. Idaho Falls, ID: Idaho Operations Office, U.S. Atomic Energy Commission, IDO­12035, June 1964.

Hawley, Jr., C.A., Editor. Controlled Environmental Radioiodine Tests at the National Reactor Testing Station 1965 Progress Report. Idaho Falls, ID: Idaho Operations Office, U.S. Atomic Energy Commission, IDO­12047, February 1966.  "

Lawrence Berkeley Laboratory

rkeley Laboratory
LBL-1. Treatment of Leukemia with Phosphorus-32

BETWEEN 1936 AND 1947, patients with various types of leukemia were treated with phosphorus­32 (P³²) with and without supplemental x­ray treatments. Approximately 129 patients with chronic myelogenous leukemia and 100 patients with chronic lymphatic leukemia were treated at the Radiation Laboratory and the Donner Laboratory of the University of California in Berkeley and San Francisco. Previously it had been demonstrated that radiophosphorus concentrated in the bone marrow and soft tissue of leukemic mice. Therefore, it was expected that P³² would provide a highly localized radiation source for human leukemic patients.

Patients were administered 1 to 2 millicuries of P³² per week for 4 to 8 weeks, although higher doses were also included. Approximately half of the patients studied had previously received x­ray treatment.

It was found that P³² treatment increased the quality of life for chronic myelogenous leukemic patients, but did not prolong the duration of life. In the case of chronic lymphatic leukemia patients, the quality of life was improved and the duration was prolonged. Based on these findings, an unspecified number of chronic lymphatic patients were treated with P³² through 1960. This research was partly supported by grants from the International Cancer Research Foundation. (Included in The DOE Roadmap of February 1995)

References

Lawrence, J.H., R.L. Dobson, B.V.A. Low­Beer, and B.R. Brown. AChronic Myelogenous Leukemia.@Journal of the American Medical Association. Vol. 136, 1948, pp. 672B677.

Lawrence, J.H., B.V.A. Low­Beer, and J.W.J. Carpender. AChronic Lymphatic Leukemia.@Journal of the American Medical Association. Vol. 140, 1949, pp. 585B588. "

LBL-2. Metabolic Studies of Bone Tumors Using Strontium-89

AN EXPERIMENT WAS conducted in 1942 at the Radiation Laboratory of the University of California, Berkeley on the uptake of radiostrontium by bone tumors. Strontium­89 (Sr⁸⁹) was administered to six subjects prior to biopsy or amputation. Tissue samples were collected and analyzed to determine the Sr⁸⁹ uptake.

The subjects consisted of five males and one female, ranging in age from 9 to 54 years. Five of the subjects received intravenous injection solutions which ranged from 326 to 1,462 microcuries. The sixth subject was given 1,183 microcuries of Sr⁸⁹ orally.

This experiment showed that Sr⁸⁹ had therapeutic value in treating certain types of bone cancers. Some of these cancer patients also received therapeutic amounts of Sr⁸⁹ (a few millicuries), but details are not available. This research was supported by the Rockefeller Foundation and the Columbia Fund for Medical Physics. (Included in The DOE Roadmap of February 1995)

References

Treadwell, A. de G., B.V.A. Low­Beer, H L. Friedell, and J.H. Lawrence. AMetabolic Studies on Neoplasm of Bone with the Aid of Radioactive Strontium.@American Journal of the Medical Sciences. Vol. 204, 1942, pp. 521B523.  "

LBL-3. Inhalation Studies Using Carbon-11

FROM 1944 TO 1945, the Aero Medical Laboratory, University of California, Berkeley and the Department of Physiology, Columbia University conducted a collaborative study using carbon-11 (C¹¹). The radioactive carbon was used as a tracer to determine whether carbon monoxide (CO) oxidizes to carbon dioxide (CO₂) in the human body.

The subjects consisted of four men, including three of the researchers conducting the experiment. The four men inhaled a relatively large amount of CO labeled with C¹¹, after which they breathed oxygen. During this time, their expired CO₂ was collected and measured for C¹¹, the presence of which would prove that the human body could convert CO to CO₂. Geiger counters were placed over various parts of the body (thigh, chest, spleen, and liver) to measure the uptake and elimination of CO₂. The CO oxidized to CO₂ amounted to less than 0.1 percent of the CO lost from the blood. This work was supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Lawrence, J.H. Positron Emitting Isotopes: Investigative and Diagnostic Studies. Berkeley, CA: Lawrence Berkeley Laboratory, John Hundale Lawrence Files, pp. 247B262. Accession 434­92­0066, File Code 19­14­6, Carton 15, Folder APositron Emitting Isotopes.A

Tobias C.A., J.H. Lawrence, F.J.W. Roughton, W.S. Root, and M.I. Gregersen. AThe Elimination of Carbon Monoxide from the Human Body with Reference to the Possible Conversion of CO to CO₂.@American Journal of Physiology. Vol. 145, No. 2, December 1945, pp. 253B263.  "

LBL-4. Inhalation of Zirconium-89 on Smoke Particles

INHALATION STUDIES were conducted at Lawrence Berkeley Laboratory in approximately 1945 using an active smoke containing zirconium­89 (Zr⁸⁹). One member of the research team was the only subject. The purpose of this experiment was to determine the degree of retention by the lungs of very finely divided active smoke suspended in air. The results showed that almost 100 percent of the inhaled activity (about 0.5 microcurie of Zr⁸⁹) was retained within the lungs and upper respiratory tract. This work was supported by the U.S. Atomic Energy Commission. (Previously described in #30 on the original list of 48 experiments released by DOE in June 1994 and included in The DOE Roadmap of February 1995)

References

Scott, K.G., D. Axelrod, J. Crowley, and J.G. Hamilton. ADeposition and Fate of Plutonium, Uranium and Their Fission Products Inhaled as Aerosols by Rats and Man.@Archives of Pathology. Vol. 48, No. 1, JulyBDecember 1949, pp. 31B54.  "

LBL-5. Radioactive Phosphorus as a Possible Diagnostic Procedure for Breast Tumors

IN 1946 , the University of California Hospital, San Francisco, employed phosphorus­32 (P³²) in tracer studies to develop a new diagnostic procedure for distinguishing between malignant and benign breast tumors. Twenty­five female patients with breast tumors were included in the study. All patients had been scheduled for surgery.

Each patient was intravenously administered 300 to 500 microcuries of P³² as sodium phosphate 24 or 48 hours prior to surgery. Surface measurements were made over the tumor and over a control area on the opposite normal breast, 2, 4, 6, and 20 hours after the injection of P³². An increase in counts was found over the surface of malignant tumors, whereas counts were not elevated over benign tumors. The malignancy of the tumor was determined after surgical removal.

Results indicated that P³² might be used as a diagnostic procedure for breast cancer, except for very slow­growing or deep­seated cancers. (Included in The DOE Roadmap of February 1995)

References

Low­Beer, B.V.A., H.G. Bell, H.J. McCorkle, R.S. Stone, H.L. Steinbach, and W.B. Hill. AMeasurement of Radioactive Phosphorus in Breast Tumors in Situ: A Possible Diagnostic Procedure.@Radiology. Vol. 47, pp. 429B496.  "

LBL-6. Comparison of the Uptake of Zirconium-95 in Tumor and Normal Tissue

IN 1946, at the University of California, San Francisco and the Crocker Radiation Laboratory, University of California, Berkeley research was carried out to study the uptake and deposition of zirconium. The subject, a 55­year­old female patient with a reticulo endothelial tumor that had arisen in the spleen and then metastasized to the liver and left leg, was given a test dose of zirconium-95 (Zr⁹⁵).

The subject was administered 1.76 millicuries of Zr⁹⁵ in saline by intravenous injection 24 hours prior to a scheduled midthigh amputation of the left leg. Samples of the tumor, as well as normal tissue, were later obtained from the limb for Zr⁹⁵ assay.

The tumor was found to have greater uptake of Zr⁹⁵ than the normal tissues of the body. External counting 2 hours after the Zr⁹⁵ injection showed that the liver contained about 90 percent of the total measurable deposition and the tumor had about 10 percent of the total deposition. This study was supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Low­Beer, B.V.A., K.G. Scott, J.G. Hamilton, and R.S. Stone. AComparative Deposition of Zr⁹⁵ in a Reticulo Endothelial Tumor to Normal Tissues in a Human Patient.@Berkeley, CA: University of California Radiation Laboratory, UCRL­68.   "

LBL-7. Autoradiographic Studies of the Distribution of Radiolabeled Lewisite and Mustard Gas on Skin

THIS EXPERIMENT was conducted in 1947 at the Crocker Radiation Laboratory, University of California, Berkeley and the University of California Medical School in San Francisco. The experiment sought to determine the distribution of mustard and lewisite in skin and eye tissues. These two chemical­warfare gases were labeled with radioactive sulfur (S³⁵) and radioactive arsenic (As⁷⁴). Small areas of the skin of four normal subjects were exposed to the two labeled gases.

Two experiments were performed with mustard gas labeled with S³⁵. The first involved a 10­minute exposure to 475 micrograms of labeled chemical; the second, a 15­minute exposure to 475 micrograms. In both cases, the exposed area was 0.43 square centimeter and biopsy specimens of these areas were taken 24 hours after exposure.

Two experiments were also performed on lewisite labeled with 10 micrograms of As⁷⁴; the first involved a 10­minute exposure to 475 micrograms of lewisite; the second, a 15­minute exposure to 475 micrograms. The new technique of autoradiography was used to determine the skin layer at which the fixation took place on the biopsied human skin samples. Lewisite was found to fix primarily in the epidermis, and mustard gas fixed in both the epidermis and dermis. (Included in The DOE Roadmap of February 1995)

References

Axelrod, D.J., and J.G. Hamilton. ARadio­Autograph Studies of the Distribution of Lewisite and Mustard Gas in Skin and Eye Tissues.@American Journal of Pathology. Vol. 23, 1947, pp. 389B411.  "

LBL-8. Injection of Americium-241

ON JUNE 10, 1947, at the University of California, San Francisco a 16­year­old Chinese male patient at the Chinese Hospital in San Francisco, identified as Cal­A, with osteogenic sarcoma of the left femur and general metastases, received an intramuscular injection of americium­241 (Am²⁴¹). The estimated activity administered was about 0.2 microcurie. The same day, two rats were given intramuscular injections of 1 milliliter of solution made from the same specifications as the Cal­A injection.

Readings of the subject=s urine and feces were collected through at least June 24, 1947. On June 12, 1947, the subject=s leg was amputated at the left midthigh. Samples of the amputation tissue were dissected the next day. The samples were measured for isotope uptake, as the tumor was expected to have higher uptake than normal body tissues. Studies were made of the tumor; the bone tissue in which the tumor was found; the surrounding tissues, both bone and connective; and the muscles. Measurements from the amputated tissues were compared with the rat data; the patient was discharged on July 27, 1947.

Rat data showed considerable uptake by the liver; human data appeared to show 13 to 20 percent uptake by the bone. The patient died of preexisting ailments on June 15, 1948.

The experiment appears to have been done as a comparison to previous human studies involving plutonium, as data sheets for Cal­A show standards for measurements set against Cal­1 (a human injected with plutonium­238). (Included in The DOE Roadmap of February 1995)

References

Lawrence Berkeley Laboratory, Joseph G. Hamilton Records, Archives and Records Office, Folder Am H (95H).  "

LBL-9. Uptake of Iodine-131 in Thyroids of Psychiatric Patients

FROM JULY 1949 TO APRIL 1950 a cooperative research project was conducted by the Departments of Psychiatry, Radiology, and Medicine at the University of California Medical School and the Langley Porter Clinic in San Francisco. The objective of this project was to determine whether thyroid function was normal or abnormal in persons with mental illness. Sixty­five subjects were selected from the regular in­patient group at the Langley Porter Clinic.

Among the subjects were patients with schizophrenia, manic­depression, mixed psychoneurosis, and anorexia nervosa. A control group was selected of volunteers from the clinic=s, clerical, and medical staff.

Subjects were injected with 150 microcuries of iodine­131 (I¹³¹); subsequently, the concentration of I¹³¹ in the thyroid was then measured six times over a 72-hour period. The test and control groups underwent medical and psychiatric evaluations, including serum­bound iodine, basal metabolism, plasma cholesterol, and electroencephalogram.

No abnormal thyroid function was found in the group with mental illness and no significant differences were detected between the patients and the controls in this study. This study was partly funded by the U.S. Atomic Energy Commission. (Previously described in #2 on the original list of 48 experiments released by DOE in June 1994 and included in The DOE Roadmap of February 1995)

References

Bowman, K.M., E.R. Miller, M.E. Dailey, A. Simon, B. Frankel, and G.W. Lowe. AThyroid Function in Mental Disease Measured with Radioactive Iodine, I¹³¹.@The American Journal of Psychiatry. Vol. 106, No. 7, February 1950.

Stone, R.S. Biological Effects of Radiations from External and Internal Sources, Progress Report July 1, 1949 to April 15, 1950. San Francisco: University of California Radiation Laboratory, April 1950. U.S. Department of Energy Archives, Record Group 326, U.S. Atomic Energy Commission, Box 3358, Folder 22.  "

LBL-10. Sodium-24 Uptake Studies on Patients with Rheumatoid Arthritis

DURING THE MID-1940s through the early 1950s, Lawrence Berkeley Laboratory conducted studies on the uptake of sodium­24 (Na²⁴) to evaluate vascular abnormalities in persons with rheumatoid arthritis.

Sodium­24 was administered by intravenous injection, usually in 50­microcurie amounts. Systemic transport of Na²⁴ was followed, using two gamma counters: one in the subject=s hand, the other placed under a knee.

The results showed an impeded blood flow in diseased areas of the body. Uptake of Na²⁴ in the knee joint was also studied after three patients drank a solution of sodium chloride labeled with Na²⁴ in water. (Included in The DOE Roadmap of February 1995)

References

Tobias, C. Sodium Uptake Studies. Berkeley, CA: Lawrence Berkeley Laboratory, Cornelius A. Tobias Papers, Accession 434­89­100, File Code 10­08­063, Carton 25/38, Folder ASodium Uptake Studies.@ "

LBL-11. Blood Volume Studies with Iron-59, Phosphorus-32, and Chromium-51 Involving Inmates at San Quentin Prison

FROM 1949 TO THE LATE 1950s, the University of California conducted studies involving radioactive isotopes using inmates at San Quentin Prison as volunteer subjects. Studies included the following: (a) 1949 to 1951: studies on red blood-cell productionCBlood was drawn from participants, labeled with iron­59 (Fe⁵⁹), and reinjected into the respective subjects. Four samples were drawn at specific intervals over the next 2 hours. The procedure was repeated for 4 successive days, during which Fe⁵⁹­labeled red blood cells were counted. (b) 1950: studies on blood volumeCAt least 13 participants had blood drawn, labeled with phosphorus­32 (P³²), and reinjected. Blood volume in the subjects was subsequently measured. (c) late 1950s: Studies on red cell volumeCChromium­51 (Cr⁵¹) was used as a label to measure red blood-cell volume in 201 healthy participants. (Included in The DOE Roadmap of February 1995)

References

Wennesland, R., E. Brown, J. Hopper, Jr., J.L. Hodges, Jr., O.E. Guttentag, K.G. Scott, I.N. Tucker, and B. Bradley. ARed Cell, Plasma, and Blood Volume in Healthy Men Measured by Radiochromium (Cr⁵¹) Cell Tagging and Hemocrit.@The Journal of Clinical Investigation. Vol. 38, No. 7, July 1959, pp. 1,065B1,077.

Donner Laboratory Clinical Books, 1946B1977. Patient Sheets from February to March 1950, noted ASan Quentin@after patient=s name. Lawrence Berkeley Laboratory. Donner Laboratory Clinical Logs and Notebooks, Accession 439­89­0151, File Code 8­2­2, Carton 7/10, Binder 2.

Letter. J.H. Lawrence to J.H. Corley. August 17, 1949. Lawrence Berkeley Laboratory, Administrative Files of Administrative Assistants to the Directors of the Biology and Medicine Division and Donner Laboratory, Accession 434­90-0209, File Code 16­5­22, Carton 2, Folder AHistorical Donner Laboratory.@  "

LBL-12. Blood and Tissue Studies with Iron­59

THIS RESEARCH was conducted at the Donner Laboratory, University of California, Berkeley in the early 1950s. The purpose of this study was to investigate the rates and pathways of iron transport in the human body, including the differences in iron turnover rates between normal individuals and patients with anemia. The subjects consisted of 22 individuals with anemia and other diseases and 16 normal individuals.

Between 5 and 30 microcuries of radioactive iron-59 (Fe⁵⁹) globulin were injected intravenously to label the circulating plasma iron globulin. External radiation measurements were made on the liver, spleen, and bone marrow using a gamma­fluorescence detector. In addition, plasma and whole-blood samples were analyzed for Fe⁵⁹ content.

The results showed that iron turnover rates varied, the exact rate depending on the disease state of the patient. This research was partly funded by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Elmlinger P.J., R.L. Huff, C.A. Tobias, and J.H. Lawrence. AIron Turnover Abnormalities in Patients Having Anemia: Serial Blood and In Vivo Tissue Studies with Fe⁵⁹.@Acta Haematologica. Vol. 9, No. 2, February 1953, pp. 73B96.

Huff, R.L., C.A. Tobias, and J.H. Lawrence. AA Test for Red Cell Production.@Acta Haematologica. Vol. 7, No. 3, March 1952, pp. 129B143.  "

LBL-13.

(Duplicate of LBL-10 in The DOE Roadmap)

LBL-14. Studies on the Rate of Uptake of Iodine-131 in the Thyroid

IN THE EARLY 1950s, studies were conducted at the University of California, San Francisco on various aspects of thyroid function in patients with normal and abnormal thyroid glands. The 427 study participants included patients with normal and abnormal thyroids as well as goiters or uncertain thyroid functions.

After the subjects drank a solution containing approximately 100 microcuries of iodine­131 (I¹³¹), an external gamma counter was placed over the thyroid to measure the uptake of radioiodine. A good correlation was found between high rates of uptake and hyperthyroidism, and between lower rates and absence of hyperthyroidism.

Further research was also conducted to study aspects of the physiology of the thyroid and other endocrine glands. Studies were conducted on obese patients, and on adult and child hyperthyroid patients requiring thyroid stimulating hormone. This research was funded by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Miller, E.R., M.E. Dailey, A.V. Holmes, G.L. Alexander, and G.E. Sheline. AStudies with Radioiodine: I. Function and Rate of I¹³¹ Uptake of Thyroid.@Radiology. Vol. 57, No.1, July 1951, pp. 37B47.

Annual Report of Cancer Activities of the Radiological Laboratory. San Francisco: University of California School of Medicine, 1953, pp. 1B9. Special Collections, The Library, University of California, San Francisco.  "

LBL-15. Measures of Body Fat and Related Factors in Normal Adults Using Potassium-40, Cesium-137, and Tritium

DURING 1950 TO 1960,  the Donner Laboratory at the University of California, Berkeley, in collaboration with the California State Department of Public Health and the Bureau of Public Health Nutrition, conducted a series of experiments using natural potassium­40 (K⁴⁰) and cesium­137 (Cs¹³⁷) to measure body composition. The experiments were designed to accurately determine the human body=s total water content, body fat, protein content, and bone mineral content. In all, 2,301 healthy volunteers were used for these experiments.

The laboratory analyses included measurements of total-body water after an oral tracer dose of tritium, analysis of specific gravity by the helium dilution technique, and whole-body counting of K⁴⁰. This study was partly supported by a grant from the National Institutes of Health. (Included in The DOE Roadmap of February 1995)

References

Steinkamp, R.C., N.L. Cohen, W.R. Gaffey, T. McKey, G. Bron, W.E. Siri, T.W. Sargent, and E. Isaacs. AMeasures of Body Fat and Related Factors in Normal Adults­II.@Journal of Chronic Diseases. Vol. 18, 1965, pp. 1,279B1,289. "

LBL-16. Study of Ascitic Fluid Using Tritium-Labeled Water and Phosphorus-32

DURING 1951 AND 1952, the University of California Donner Laboratory and the Highland Alameda County Hospital, Oakland conducted experiments to determine the total amount of ascitic fluids in humans. Tritium (H³) was used to trace the flow of water into, and out from, the peritoneal cavity.

Six patients with ascites (a condition characterized by fluid buildup in the peritoneal cavity) were injected with 2 microcuries of tritium­labeled water, either intravenously or intraperitoneally. Over the following 7 to 24 hours, samples of blood and ascitic fluid were taken. Blood samples were labeled with phosphorus­32 (P³²) and 1 milliliter of the labeled blood was injected into the peritoneal cavity.

This study showed that the water content of ascitic fluid entered and left the peritoneal cavity at a very rapid rate. It also showed that the peritoneal surfaces of both normal and diseased subjects reabsorbed large volumes of fluid. This work was supported by the Life Insurance Medical Research Fund and the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Prentice, T.C., W. Siri, and E.E. Jones. AQuantitative Studies of Ascitic Fluid Circulation with Tritium­Labeled Water.@American Journal of Medicine. Vol. 13, No. 6, December 1952, pp. 668B673.  "

LBL-17. A Physiological Study in the Peruvian Andes Using Iron-59

THE DONNER LABORATORY of Medical Physics, University of California, Berkeley used iron­59 (Fe⁵⁹) in high­altitude studies similar to the previously conducted studies using tritium (H³). The purpose of these experiments was to investigate the physiology of reduced barometric pressure, particularly as seen in high­altitude flights, and the physiology and treatment of various hematopoietic (blood­forming) disorders, especially polycythemia rubra vera, leukemia, and aplastic anemia.

In these studies, reported in 1952, healthy subjects (medical students from the University of San Marcos, Lima, Peru) and native Peruvians in the Andes mountains were studied. Four Andean natives suffering from pulmonary silicosis (as well as high­altitude polycythemia rubra vera) were also studied.

A few micrograms of Fe⁵⁹ were incubated for 20 minutes with 10 to 20 milliliters of the subject=s plasma and then injected into the subjects. After injection, Fe⁵⁹ analysis was made on plasma samples taken at hourly intervals for 4 to 5 hours. Acclimatization to high altitude was found to be related to changes in blood volume, plasma volume, and red blood­cell mass. Postexposure plasma­iron turnover rates and red cell renewal rates increased to roughly twice their normal values in less than 12 hours at high altitude. This study was supported by the U.S. Navy, the U.S. Air Force, and the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Lawrence, J.H., R.L. Huff, W. Siri, L.R. Wasserman, and T.G. Hennessy. AA Physiological Study in the Peruvian Andes.@Acta Medica Scandinavica. Vol. 142, No. 2, 1952, pp.117B133.  "

LBL-18. Studies on the Metabolism of Glycine Labeled with Carbon-14

DURING THE EARLY 1950s, researchers at the University of California Donner Laboratory conducted studies on human carbon metabolism using carbon-14 (C¹⁴). Twelve patients participated in two studies that examined the distribution, retention, and excretion of C¹⁴.

In one study, four patients, ranging in age from 29 to 52 years, were intravenously administered 100 microcuries of C¹⁴-labeled glycine (an essential nutrient and a dietary supplement). Activity as C¹⁴O₂ was measured in their exhaled breath and in tissue samples subsequently collected at autopsy. Five additional patients received similar injections and C¹⁴ activity was measured in their urine samples.

In a second study, which examined the retention of C¹⁴ in hemoglobin precursors, three patients were administered 100 microcuries of C¹⁴-labeled glycine and activity was measured in hippuric acid isolated from their urine samples.

The results of these studies showed that exhalation of C¹⁴O₂ was the primary route of C¹⁴ excretion, with a small percentage being excreted through urine. The studies also showed a small concentration in tissue and a biological retention half-time for C¹⁴ of 50 days. Finally, the studies showed that retention in the blood was due to absorption by newly formed cells, not long-lived existing cells. This work was supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995, and since revised)

References

Berlin, N.I., B.M. Tolbert, and J.H. Lawrence. AStudies in Glycine-2-C¹⁴ Metabolism in Man: I. The Pulmonary Excretion of C¹⁴O₂.@Journal of Clinical Investigation. Vol. 30, 1951, pp. 73B77.

Berlin, N.I., B.M. Tolbert, and J.H. Lawrence. AStudies in Glycine-2-C¹⁴ Metabolism in Man: II. Tissue Distribution.@Journal of Clinical Investigation. Vol. 31, 1952, pp. 335B337.

Berlin, N.I., B.M. Tolbert, and H.C. Lee. AStudies in Glycine-2-C¹⁴ Metabolism in Man: III. The Urinary Excretion of C¹⁴ and Cumulative Radiation Dosimetry.@Journal of Clinical Investigation. Vol. 32, 1953, pp. 1B4.

Berlin, N.I., C. Hewitt, and C. Lotz. AHippuric Acid Synthesis in Man After the Administration of ["-¹⁴C] Glycine.@The Biochemical Journal. Vol. 58, 1954, pp. 498B503.  "

LBL-19. Astatine-211 and the Thyroid

THE OBJECTIVE of this experiment was to test the uptake of astatine­211 (At²¹¹) and to evaluate its potential benefits in the treatment of thyroid diseases. Eight subjects were injected with 50 microcuries of the 7­hour half­life alpha emitter At²¹¹. These experiments were conducted at the University of California Hospital during early 1954. (Previously described in #37 on the original list of 48 experiments released by DOE in June 1994 and included in The DOE Roadmap of February 1995)

References

Hamilton, J.G., P.W. Durbin, and M.W. Parrott. AAccumulation of Astatine by Thyroid Gland in Man.@In Proceedings of the Society for Experimental Biology and Medicine. Vol. 86, 1954, pp. 366B369.

Hamilton, J.G., P.W. Durbin, and M.W. Parrott. AComparison of Acute and Chronic Changes Produced in Rats by I¹³¹ and At²¹¹ at Lethal Levels. Preliminary Data on the Uptake of At²¹¹ in Patients with Thyroid Disease.@Chapter 24 in Proceedings of the 2nd Radioisotope Conference in Oxford, England. pp. 219B231. London: Butterworth Scientific Publications, July 1954.  "

LBL-20. Body Water at Sea Level and at High Altitudes by Tritium Analysis

IN 1954, SCIENTISTS from the Donner Laboratory, University of California, Berkeley, and the Instituto de Biologia Andina, Lima, Peru, used tritium (H³) to determine changes in weight and total-body water for subjects living in Lima at high altitudes and at sea level. Two groups of subjects were studied. The first group consisted of 15 young male medical students; the second group consisted of 13 normal male Peruvian Indian mine workers.

The tritium was administered both orally and intravenously. The mean values of body water for the two groups was normal for their age range and occupations. This research was supported by the U.S. Public Health Service, the U.S. Atomic Energy Commission, and the U.S. Air Force. (Included in The DOE Roadmap of February 1995)

References

Siri, W.E., C. Reynafarje, N.I. Berlin, and J.H. Lawrence. ABody Water at Sea Level and at Altitude.@The Journal of Applied Physiology. Vol. 7, No. 3, November 1954, pp. 333B334.  "

LBL-21. High-Energy-Beam Irradiation of Breast Cancer Patients

DURING THE 1950s AND EARLY 1960s, researchers at Donner Laboratory and Lawrence Radiation Laboratory, University of California conducted studies on therapeutic heavy particle beam irradiation. The purpose of these studies was to evaluate the results of pituitary irradiation in the treatment of a variety of hormone-dependent diseases.

An early study involved 26 breast cancer patients ranging in age from 27 to 70 years who received between 14,000 and 30,000 rads of 340-MeV (million electron-volt) radiation to the pituitary gland over periods of 9 to 63 days. Beginning in 1954, 159 breast cancer patients received therapy involving 900-MeV alpha particles produced by an accelerator. All patients were either in a terminal state or a state of rapid disease advancement despite all routinely available treatment. Radiation doses to the pituitary ranged from 14,000 to 17,000 rads delivered over 11 days. During the course of treatment, patients also underwent various diagnostic tests involving the use of internally administered iodine-131, carbon-14, and calcium-47. The results of these studies showed limited supression of tumor growth.

Beginning around 1956, 79 diabetes patients received heavy particle therapy in an attempt to slow degeneration of the retina. Doses ranged from 8,000 to 12,000 rads delivered over 11 days. The results of the study showed that diabetics with advanced complications did not benefit much from this therapy. Also beginning around 1956, 21 patients received therapy for acromegaly (a disease characterized by enlargement of the head, hands, feet, and thorax). Doses ranged from 3,000 to 7,200 rads delivered over 11 days. Follow-up studies showed beneficial changes in patients.

From 1957 to 1961, three female patients with Cushing=s disease received therapy of 5,000 to 10,000 rads delivered over 11 days. All showed some benefit, from minor improvement to general remission. Three patients received up to 10,000 rads over 11 days to treat malignant exophthalmos. One patient showed beneficial results.

In 1958, one breast cancer patient was treated with direct tumor therapy (not pituitary irradiation) of 2,500 rads over 6 days and experienced temporary remission. These studies were supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995, and since revised)

References

Lawrence, J.H., C.A. Tobias, J.L. Born, A. Gottschalk, J.A. Linfoot, and R.P. Kling. AAlpha Particle and Proton Beams in Therapy.@Journal of the American Medical Association. Vol. 186, No. 3, October 1963, pp. 236B245.

Tobias, C.A., J.H. Lawrence, J.L. Born, R.K. McCombs, J.E. Roberts, H.O. Anger, B.V.A. Low-Beer, and C.B. Huggins. APituitary Irradiation with High-Energy Proton Beams: A Preliminary Report.@Cancer Research. Vol. 18, No. 2, February 1958, pp. 121B134.  "

LBL-22. Iron-59 Metabolism in Patients with Cancer and Anemic Conditions

STUDIES WERE CONDUCTED in 1959 at the University of California Lawrence Radiation Laboratory on the metabolism of iron in humans using iron­59 (Fe⁵⁹) as a tracer. The aim of these studies was to determine the effects of age, gender, and health status on iron metabolism in humans. Approximately 80 cancer patients and subjects with various anemias, hemochromatosis (a disease characterized by an excessive absorption of iron), and iron deficiencies were used in these studies.

The rate of hemoglobin synthesis, mean red blood­cell life span, and mean time required for hemoglobin formation within the total red cell volume were measured. Gastrointestinal bleeding was correlated with iron and red cell movement in seven subjects. This work was supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Polycove, M., and J.H. Lawrence. AIron Metabolism.@University of California Lawrence Radiation Laboratory Project Description. Berkeley, CA: Lawrence Berkeley Laboratory, June 30, 1959. Cornelius A. Tobias Papers, Accession 434­92­0154, File Code 19­14­43, Carton 21, Folder AProgram Book.@ "

LBL-23. Radionuclide Studies to Determine Bone Marrow Distribution in Humans

IN THE EARLY 1960s, at the Donner Laboratory and the Lawrence Radiation Laboratory, University of California, Berkeley, iron­52 (Fe⁵²), iron­59 (Fe⁵⁹) and technetium­99m (Tc^99m)Bsulfur colloid were administered to study marrow distribution. The marrow, liver, and spleen were then imaged, using conventional scanners or scintillation cameras. Administered activities ranged from 3 to 100 microcuries. Samples of bone marrow, plasma, red cells, and liver were analyzed to determine tissue activity over time. Subjects included hospital patients and normal volunteers, including children. This work was supported in part by the U.S. Atomic Energy Commission and in part by a grant from the National Cancer Institute of the National Institutes of Health. (Included in The DOE Roadmap of February 1995)

References

Lawrence, J.H. Positron Emitting Isotopes: Investigative and Diagnostic Studies. Berkeley, CA: Lawrence Berkeley Laboratory, pp. 247B262. John Hundale Lawrence Files, Accession 434­92­0066, File Code 19­14­6, Carton 15, Folder APositron Emitting Isotopes.@

Van Dyke D.C., H.O. Anger, and Y. Yano. AProgress in Determining Bone Marrow Distribution In Vivo.@Progress in Atomic Medicine. Vol. 2, 1968, pp. 65B84.  "

LBL-24. Iron Kinetics and Hemoglobin Synthesis in Human Subjects with Iron-59BBound Plasma

THIS STUDY WAS conducted in approximately 1959 at the University of California, Berkeley in collaboration with the Veterans Administration in Boston. Its purpose was to develop a suitable mathematical model of hemoglobin synthesis, using sequential measurements of iron­59 present in human plasma, red cells, and peripheral blood. Data were obtained from 13 normal, healthy subjects (1 female and 12 male volunteers) between the ages of 24 and 72 years, and 6 male hospital patients with endogenous hemochromatosis.

Five to 20 milliliters of plasma labeled with 10 to 40 microcuries of iron­59 (Fe⁵⁹) were intravenously injected into the subjects. Plasma and erythrocyte radioactivity were measured with a scintillation counter. This study was supported by the U.S. Atomic Energy Commission with partial support by a grant from the U.S. Public Heath Service. (Included in The DOE Roadmap of February 1995)

References

Pollycove, M., and R. Mortimer. AThe Quantitative Determination of Iron Kinetics and Hemoglobin Synthesis in Human Subjects.@Journal of Clinical Investigation. Vol. 40, 1961, pp. 753B772.   "

LBL-25. Intestinal Iron Absorption Studies Using Iron-52, Iron-55, and Iron-59

IN A STUDY CONDUCTED at the Donner Laboratory, University of California, Berkeley in 1966, radioactive isotopes of iron were used to measure the rate of iron absorption into the plasma and its distribution in the gastrointestinal tract.

Forty microcuries of iron­52 (Fe⁵²) were administered orally to six fasting normal subjects. Just prior to the oral dose, iron turnover studies were performed using 2 microcuries of transferrin­bound iron­59 (Fe⁵⁹) injected intravenously; the subjects were then whole-body counted. For the iron turnover studies, 20 to 30 microcuries of iron­55 (Fe⁵⁵) were injected into the same subjects. Photoscans of the abdomen using the Anger positron camera were taken throughout the study. The maximum rate of intestinal iron absorption was found to occur at the time when iron was in the upper gastrointestinal tract. This work was supported by the U.S. Atomic Energy Commission. (Included in The DOE Roadmap of February 1995)

References

Fawwaz, R.A., H.S. Winchell, M. Pollycove, T. Sargent, H. Anger, and J.H. Lawrence. AIntestinal Iron Absorption Studies Using Iron­52 and Anger Positron Camera.@Journal of Nuclear Medicine. Vol. 7, 1966, pp. 569B576.  "

LBL-26. Chromium-51 Metabolism Studies in Patients with Hemochromatosis

CHROMIUM-51 METABOLISM studies were conducted at Lawrence Berkeley Laboratory on healthy males and on patients with hemochromatosis (a disease characterized by an excessive absorption of iron).

Five normal male subjects were injected with 100 microcuries of chromium-51 (Cr⁵¹) to study the retention of chromium. This study was conducted to show that hemochromatic diabetes was due to the exclusion of chromium from either the carrying agent or from the liver because of saturation by iron.

Eleven subjects were injected with Cr⁵¹­chloride. Among the subjects were patients with varying degrees of hemochromatosis, including two hemochromatotic patients depleted of excess iron and two subjects with excess iron but no clinical disease. All of the subjects were followed by whole-body counting for up to 6 months.

The results showed that the exclusion of chromium occurs principally at binding sites in the liver. Two further studies were conducted on chromium metabolism using plasma analysis, Cr⁵¹ clearance rates, the whole-body scanner, and the whole-body counter. This work was supported by the U.S. Department of Energy. (Included in The DOE Roadmap of February 1995)

References

Lim, T.H., T. Sargent, and N. Kusubov. AKinetics of Trace Element Chromium (III) in the Human Body.@American Journal of Physiology. Vol. 244, Vol. 4, April 1983, pp. R445B454.

Sargent T.W., and H.H. Stauffer. AHuman In Vivo Kinetics of Radionuclide Trace Metals in Health and Disease.@University of California Lawrence Berkeley Laboratory Project Description. Berkeley, CA: Lawrence Berkeley Laboratory, May 1, 1976. Lawrence Berkeley Laboratory, Cornelius A. Tobias Papers, Accession 434­32­0154, File Code 19­14­43, Carton 22, UCLBL Project Descriptions.  "

LBL-27. Calcium-47 Retention Studies in Juvenile Diabetics

THIS RESEARCH was conducted at the Berkeley Donner Laboratory in the early 1970s. This study was undertaken to determine the rate of uptake and retention of calcium­47 (Ca⁴⁷) in juvenile diabetics. The subjects consisted of eight healthy individuals, of various ages and diets, and three juvenile diabetics (ages 23, 26, and 26).

One to 25 microcuries of Ca⁴⁷ was intravenously administered and the retention of Ca⁴⁷ in the whole body was determined by direct in vivo counting.

The whole-body retention of Ca⁴⁷ did not significantly vary over the wide range of calcium and protein intakes and ages of healthy subjects. Diabetics excreted Ca⁴⁷ at a higher rate.

This work showed a decreased rate of bone mineralization in diabetics. The research was supported by the U.S. Energy Research and Development Administration. (Included in The DOE Roadmap of February 1995)

References

Sargent, T., J. Linfoot, and H. Stauffer. ADecreased Whole-Body Retention of Ca⁴⁷ in Juvenile Diabetics.@IRCS Medical Science Journal. Vol. 4, 1976, p. 58.

Sargent T.W., and H.H. Stauffer. AHuman In Vivo Kinetics of Radionuclide Trace Metals in Health and Disease.@University of California Lawrence Berkeley Laboratory Project Description. Berkeley, CA: Lawrence Berkeley Laboratory, May 1, 1976. Lawrence Berkeley Laboratory, Cornelius A. Tobias Papers, Accession 434­32­0154, File Code 19­14­43, Carton 22, UCLBL Project Descriptions.  "

LBL-28. Whole-Body Counting Studies on the Retention of Copper-67 and Phosphorus-32 and Chromium-51

IN THE MID- TO LATE 1970s, the University of California Lawrence Berkeley Laboratory conducted studies on the retention of radionuclides in humans. The subjects were healthy individuals and patients with a variety of diseases.

Four subjects were injected with 100 microcuries of copper­67 (Cu⁶⁷) to determine copper uptake, retention, and excretion rates. Of the four subjects, three were healthy, and one had a copper storage disease. The results showed that there is no abnormality of total-body turnover of copper when iron stores are normal. Results also showed that for the subject with the copper storage disease, the excretion of copper was slower than for normal subjects by a factor of two.

Six subjects with diseases related to bone marrow production were injected with 1 to 5 millicuries of phosphorus­32 (P³²) to determine excretion rates. This was one of the first published studies on human whole-body phosphorus turnover.

Five subjects received injections of 100 microcuries of chromium-51 (Cr⁵¹). The whole-body retention and excretion rates of Cu⁶⁷, P³², and Cr⁵¹ were reported. This work was supported by the U.S. Department of Energy. (Included in The DOE Roadmap of February 1995 and since revised.)

References

Sargent, T.W., and H. Stauffer. AWhole-Body Counting of Retention of Cu⁶⁷, P³², and Cr⁵¹ in Man.@International Journal of Nuclear Medicine and Biology. Vol. 6, 1979, pp. 17B21.  "

LBL-29. Metabolism of Carbon-14 B Labeled Methionine in Schizophrenics

THIS RESEARCH was conducted in the 1980s at the Lawrence Berkeley Laboratory. Researchers suspected that a defect in the methyl­carbon metabolic pathway was a causative factor in schizophrenia. Methionine labeled with carbon­11 (C¹¹) or carbon­14 (C¹⁴) was administered to both schizophrenics and healthy subjects to test this hypothesis. The oxidation of methionine was studied in seven unmedicated schizophrenics, and the effect of high and low methionine in the diet was studied in control subjects. This research was supported by the National Institute of Mental Health, the Donner Laboratory, and the U.S. Department of Energy. (Included in the DOE Roadmap of February 1995)

References

Sargent, T.W. AMetabolism of [C¹⁴­Methyl]-Methionine in Schizophrenia and Affective Disorders.@U.S. Public Health Service, Food and Drug Administration, Radioactive Drug Research Committee, Report on Research Use of Radioactive Drug Study Summary. Berkeley, CA: Lawrence Berkeley Laboratory, Director=s Office, LBL Associated Laboratory, Director for Administration R&D FilesC1981B1987, Accession 434­91­0176, File Code 13­11­14, Carton 58/58, Folder ABiology/Medicine General FY87.@

Sargent, T.W. AMetabolism in Brain Disorders.@U.S. Department of Energy Field Task Proposal/Agreement. Berkeley, CA: Lawrence Berkeley Laboratory, April 1, 1982. Cornelius A. Tobias Papers, Accession 434­92­0154, File Code 19­14­43, Carton 21, Field Task Proposals/Agreements.

Sargent, T.W., N. Kusubov, S. Taylor, and T.F. Budinger. ATracer Kinetic Evidence for Abnormal Methyl Metabolism in Schizophrenia.@Biological Psychiatry. Vol. 32, 1992, pp. 1,078B1,090.   "

 ‡  ‡  ‡

LBL-30. Experimental Therapy of Multiple Myeloma with Phosphorus-32, Strontium-89, and Yttrium-90

FROM 1939 TO 1949, twenty-one male and female patients, ranging in age from 29 to 66 years, at the Donner Laboratory, University of California Radiation Laboratory, Berkeley received experimental radioisotope treatments for multiple myeloma.

Nine patients received phosphorus-32 (P³²) and strontium-89 (Sr⁸⁹); 11 received P³² alone; and 1 received colloidal yttrium-90. Levels of the administered activity differed for each patient. Subjects received 1 to 21 oral or intravenous administrations of activity, with total activity ranging from 0.3 to 103 millicuries.

The results of the treatment with P³² or Sr⁸⁹ were no better than results obtained from treatment with x­rays or stilbamidine. The Donner Laboratory was supported by the U.S. Atomic Energy Commission.

References

Lawrence, J.H., and L.R. Wasserman. AMultiple Myeloma: A Study of 24 Patients Treated with Radioactive Isotopes (P³² and Sr⁸⁹).@Annals of Internal Medicine. Vol. 33, 1950, pp. 41B55.  "

LBL-31. Experimental Therapy of Polycythemia Vera Using Phosphorus-32 and X­Rays

BETWEEN 1939 AND 1969, researchers at the Donner Laboratory, University of California Radiation Laboratory, Berkeley treated 181 polycythemia vera patients using intravenously administered phosphorus-32 (P³²) phosphate, or P³² plus x­rays. Doses usually totaled between 15 and 45 millicuries of P³² per patient. X­ray absorbed doses varied greatly between patients.

Excellent results were achieved in many cases, and the life expectancy was extended to nearly normal using this treatment. However, some of the polycythemia vera patients later developed splenic metaplasia and acute leukemia.

Follow-up studies were conducted in the late 1960s to evaluate the overall effectiveness of the therapy and to determine whether there was a relationship between therapy for polycythemia vera and the later incidence of leukemia. These studies indicated that the development of splenic myeloid metaplasia and acute leukemia are part of the evolutionary history of polycythemia vera, and that the incidence of acute leukemia in patients treated with P³² may be a result of prolonged survival rather than a direct effect of radiation dose. The Donner Laboratory was supported by the U.S. Atomic Energy Commission.

References

Lawrence, J.H. The Control of Polycythemia by Marrow Inhibition. Berkeley, CA: University of California Radiation Laboratory, Division of Medical Physics, UCRL-410, August 9, 1949.

Lawrence, J.H., H.S. Winchell, and W.G. Donald. ALeukemia in Polycythemia Vera: Relationship to Splenic Myeloid Metaplasia and Therapeutic Radiation Dose.@Annals of Internal Medicine. Vol. 70, 1969, pp. 763B771.

Wasserman, L.R., J.H. Lawrence, N.I. Berlin, R.L. Dobson, and S. Estern. AThe Bone Marrow Picture in Polycythemia Vera Before and After Treatment with Radioactive Phosphorus.@Acta Medica Scandinavica. Vol. 143, No. 6, 1952, pp. 443B449.  "

LBL-32. Studies of Iodine-131 Uptake in Hypothyroid Children

DURING THE EARLY 1940s, researchers at the University of California Medical School, San Francisco conducted tracer studies, using iodine-131 (I¹³¹) as sodium iodide as tracer, to measure the metabolism of iodine in normal and hypothyroid children.

Ten hypothyroid children and young adults ranging in age from less than 1 year to 19 years served as subjects. The amounts of I¹³¹ administered were about 25 to 50 millicuries, or what the researchers thought to be less than one-fiftieth the amount of radioiodine necessary to produce biologic changes. Uptake was measured with a Geiger counter. Six subjects were administered additional radioiodine a few months later. The results were compared with the radioiodine uptake of three normal children in a separate study.

This study showed that the thyroids of children with hypothyroidism and without goiters concentrated only small amounts of iodine compared to the thyroids of normal children or adults. This work was supported by grants from the Christine Breon Fund for Medical Research and the American Cyanamid Corporation. The principal investigator continued these studies and was supported by the Manhattan Engineer District (MED) during World War II. The University of California Medical School, San Francisco received funding from the MED through a contractual arrangement, and intermittently shared facilities and personnel with the MED-funded predecessors to LBL.

References

Hamilton, J.G., and M.H. Soley. AStudies in Iodine Metabolism of the Thyroid Gland in Situ by the Use of Radio-Iodine in Normal Subjects and in Patients with Various Types of Goiter.@American Journal of Physiology. Vol. 131, 1940, pp. 135B143.

Hamilton, J.G., M.H. Soley, W.A. Reilly, and K.B. Eichorn. ARadioactive Iodine Studies in Childhood Hypothyroidism.@American Journal of Diseases in Children. Vol. 66, 1943, pp. 495B502.

Handwritten Letter. Reilly, W.A. to J.G. Hamilton. January 4, 1949. The Personal Papers of Joseph Hamilton, Bancroft Library, The University of California, Berkeley, California.

Letter. Hamilton, J.G. to W.A. Reilly. January 10, 1950. The Personal Papers of Joseph Hamilton, Bancroft Library, The University of California, Berkeley, California.  "

LBL-33. Studies on Patients Treated with Total-Body X­Ray Irradiation

FROM 1942 TO 1946, researchers at the University of California, San Francisco conducted studies on the blood of patients at the University Hospital who had received therapeutic total-body irradiation (and directed tumor irradiation, in some cases). Patients were selected for radiation therapy by hospital staff physicians and radiation treatments were administered as part of the normal course of therapy for these patients. Only the ancillary blood studies were sponsored by the Manhattan Project.

Sixteen male and 13 female patients ranging in age from 20 to 75 years with metastatic carcinoma, lymphoma, or arthritis were studied. Patients received a series of daily fractional exposures of 5 to 50 roentgens, measured at the skin-surface entry point, with totals ranging from 100 to 300 roentgens. Patient blood samples were studied individually for up to 3 years after treatment. The purpose of this study was to determine whether the analysis of blood changes could be used to indicate occupational radiation exposures in workers on the Manhattan Project.

The studies of blood samples obtained at intervals from the treated patients showed an immediate decrease in the number of white blood cells that are formed in lymphoid tissue, followed by recovery to normal levels during the post-treatment period. Monocytes (very large white blood cells) increased in number during treatment, but returned to normal levels after treatment was completed. Significant deviations in total white blood-cell count were observed during both early and late post-treatment periods. The red blood-cell count and the concentration of hemoglobin decreased during treatment, but recovered during the early post-treatment period. A temporary anemia condition was observed in many patients during the late post-treatment period. These studies were supported by the Health Division of the Metallurgical Laboratory (a Manhattan Project site located in Chicago ).

References

Low-Beer, B.V.A., and R.S. Stone. AHematological Studies in Patients Treated By Total-Body Exposure to X­Rays.@In Industrial Medicine on the Plutonium Project, pp. 338B418. New York: McGraw-Hill, 1951.

Letter. R.S. Stone to A. Gregg. November 4, 1948.  "

LBL-34. Studies Using Radioactive Isotopes of Nitrogen, Argon, Krypton, and Xenon

BETWEEN 1943 AND 1949, the Divisions of Medical Physics and Medicine, and the Radiation Laboratory, University of California, Berkeley conducted studies on the exchange of isotopes of nitrogen, argon, krypton, and xenon gases between the body and the surrounding air. The isotopes used for this research included nitrogen-13, argon-41, krypton-79, krypton-81, krypton-85, xenon-127, xenon-133, and xenon-141. Fifteen subjects ranging in age from 17 to 26 years participated.

Subjects inhaled one of these radioactive gases for 30 to 120 minutes to receive an administered activity of about 0.4 millicurie of each isotope. The subjects were then either placed in a high-altitude chamber to simulate decompression or flown in airplanes to specific altitudes. The hands, legs, and knees of the subjects were placed into specially constructed Geiger counters to measure uptake and elimination rates of the gas isotopes. Counting of the isotopes continued for 1 hour after actual or simulated flight. Many of the subjects exercised at various Aaltitudes@in order to bring on Athe bends,@or the formation of nitrogen bubbles in the blood and tissues characterized by pain in the joints and abdomen. The subjects classified the degree of pain caused by the bends and this information was compared to the subjects=desaturation curves.

This study showed that diffusion plays very little role in the uptake of various gases by tissues, and that the rapidity of gas exchange is dependent on the ability of the blood to carry gas as well as the rate of perfusion of the tissues by blood. This work was supported by the Office of Scientific Research and Development and the Columbia Foundation. The Divisions of Medical Physics and Medicine, and the Radiation Laboratory at the University of California, Berkeley were supported by the U.S. Atomic Energy Commission.

References

Tobias, C.A., H.B. Jones, J.H. Lawrence, and J.G. Hamilton. AThe Uptake and Elimination of Krypton and Other Inert Gases by the Human Body.@The Journal of Clinical Investigation. Vol. 28, No. 6, November 1949, pp. 1,375B1,385.  "

LBL-35. Blood Oxygen Studies in Polycythemia Vera Patients and Normal Subjects Using Colloids of Zirconium-95 and Yttrium-90

IN APPROXIMATELY 1944 to 1947, scientists at the Donner Laboratory, University of California Radiation Laboratory, Berkeley conducted studies on blood oxygen saturation to learn more about the disease polycythemia vera. These studies involved measurements on 74 subjects, 48 of whom were polycythemia vera patients. Among the remainder, 18 were normal, healthy subjects, 4 had hypochromic anemia, and 3 had erythrocytosis (overproduction of red blood cells) and chronic pulmonary or cardiac disease. Blood volumes were determined for some of these subjects using colloids of zirconium-95 (Zr⁹⁵) and yttrium (probably Y⁹⁰, although not stated), or by the use of radioactively labeled red blood cells. No further details were given with regard to specific procedures or amounts of radionuclides administered as part of these studies. All patients also received chest x­rays to exclude polycythemia patients with pulmonary disease.

These studies showed that arterial blood oxygen saturation in polycythemia vera was within normal limits in resting subjects. This research was supported by the International Cancer Research Foundation, the U.S. Public Health Service, and the Sara Welt Fund. The Donner Laboratory was supported by the U.S. Atomic Energy Commission.

References

Dobson, E.L., J.W. Gofman, H.B. Jones, L.S. Kelly, and L.A. Walker. AStudies with Colloids Containing Radioisotopes of Yttrium, Zirconium, Columbium, and Lanthanum. II. The Controlled Selective Localization of Radioisotopes of Yttrium, Zirconium, and Columbium in the Bone Marrow, Liver, and Spleen.@Journal of Laboratory and Clinical Medicine. Vol. 34, No. 3, pp. 305B312.

Gofman, J.W. AStudies with Colloids Containing Radioisotopes of Yttrium, Zirconium, Columbium, and Lanthanum. I. The Chemical Principles and Methods Involved in Preparation of Colloids of Yttrium, Zirconium, Columbium, and Lanthanum.@Journal of Laboratory and Clinical Medicine. Vol. 34, No. 3, pp. 297B304.

Wasserman, L.R., R.L. Dobson, and J.H. Lawrence. ABlood Oxygen Studies in Patients with Polycythemia and in Normal Subjects. Journal of Clinical Investigation. Vol. 28, 1949, pp. 60B65.  "

LBL-36. Experimental Breast Cancer Treatment Studies Using Phosphorus-32

BETWEEN 1945 AND 1954, researchers at the University of California Medical School, San Francisco treated 89 breast cancer patients with phosphorus-32 (P³²) in combination with surgery and x­ray therapy. The treatment protocol included intravenous administration of 20 to 30 microcuries of P³² as sodium hypophosphate per kilogram of body weight, two or three times a week over 2 to 4 weeks. In addition, the treatment included local x­ray exposure up to 4,000 roentgens within 35 days. Total administered activity P³² ranged from 5.0 to 25.2 millicuries. In 1945 and 1946, eight additional breast cancer patients were treated. Treatments were suspended in 1947 because of concern regarding possible subsequent radiation-induced hematologic effects of internally deposited P³². Treatments were resumed in 1951 after no evidence of adverse effect had been observed among the previously treated patients, and an additional 81 subjects were treated over the next 4 years.

Follow-up of the treated patients through 1954 showed that 5 of the 8 patients treated during 1945 and 1946 were still alive, as were 75 of the 85 treated in 1951 or later.

The researchers concluded that further investigations and an expanded clinical experience were needed before a definitive conclusion could be reached about the success of this experimental treatment method. The University of California Medical School, San Francisco, received funding from the U.S. Atomic Energy Commission (AEC) through a contractual arrangement, and intermittently shared facilities and personnel with the AEC-funded predecessors to Lawrence Berkeley Laboratory.

References

Bell, H.G., and B.V.A. Low-Beer. ACancer of the Breast. A Preliminary Report on a New Approach to the Problem.@Journal of the American Geriatrics Society. Vol. 3, June 1955, pp. 396B398.

Low-Beer, B.V.A., and H.G. Bell. ASurgical and Radiation Treatment of Carcinoma of the Breast: A New Concept.@The American Journal of Roentgenology, Radium Therapy and Nuclear Medicine. Vol. 75, No. 6, June 1956, pp. 1,162B1,168.

Cancer Board Meeting Minutes, June 10 1953. Special Collections, The Library, University of California. Boldrey, Edwin Barkley, Papers, MSS 88-56, Carton 6. Folder ACancer Board 1953B54.@ "

LBL-37. Experimental Thyroid Cancer Treatment Using Iodine-131

BETWEEN 1945 AND 1954, researchers in the Department of Radiology, University of California Medical School, San Francisco evaluated the use of iodine-131 (I¹³¹) in the treatment of thyroid cancer. The cases of 124 patients with confirmed primary thyroid cancer were studied.

Most patients had received 1 to 2 millicuries of I¹³¹ to test their thyroid function before surgery. Of these patients, 101 showed no detectable tumor or I¹³¹ uptake after surgery. Among these subjects, 14 received 25 to 50 millicuries of I¹³¹ on one or two occasions; were then given thyroid extract; and were later checked for recurrence or development of radiation-induced effects. The 23 patients with inoperable thyroid cancer received 100 millicuries of I¹³¹. This was administered once monthly until there was no evidence of tumor tissue as indicated by the absence of I¹³¹ uptake, or until adverse effects of the treatment were observed.

The results showed that treatment of thyroid cancer with I¹³¹ often was not effective. Radiation-induced depression of all cell types in the circulating blood and bone marrow was evident to widely varying degrees, with the greatest effect being observed at the highest total I¹³¹ activity. However, the magnitude of the effect was not always correlated with the level of I¹³¹ activity. This research was funded by the U.S. Atomic Energy Commission.

References

Aggeler, P.M. The Effect of I¹³¹ On the Bone Marrow. San Francisco: University California at San Francisco, U.S. Atomic Energy Commission Report, UCSF-8. January 1954, pp. 38B57.

Alexford, A.R., and L. Berman. AThe Bone Marrow in Hyperthyroidism and Hypothyroidism.@Blood: The Journal of Hematology. Vol. 6, 1951, pp. 436B453.

Sheline, G.E., and E.R. Miller. AStudies with Radioiodine. VI. Evaluation of Radioiodine Treatment of Carcinoma of the Thyroid Based on the Experience at the University of California from 1938 to 1954.@Radiology. Vol. 69, 1957, pp. 527B545.