NUTRITIONAL BIOCHEMISTRY LABORATORIES MASSACHUSETTS INSTITUTE OF TECHNOLOGY CAMBRIDGE, MASSACHUSETTS December 19, 1945 Dr. Malcolm Farrell Fernald State School Box C Waverly 78, Massachusetts Dear Dr. Farrell: Dr. John W. Chamberlain has informed me of his preliminary conversation with you concerning the possibility of conducting a clinical study at your institution. He had requested me to give you further information as to the purposes of this investigation. About thirty years ago, it was noted that when dogs were fed a diet high in cereal (notably oats) content, they developed severe rickets. Subsequent research indicated that this effect was produced because cereals contain large amounts or phytates which form insoluble compounds with calcium that are not absorbable from the gastrointestinal tract. Thus, these cereals interfered with calcium utilization. Phytates are hexa-phosphate compounds or inositol, a substance which bus recently been shown to have vitamin activities. Chemical investigations have revealed that more than 50% of the phosphorus in cereals is combined in the form of phytates. There has been considerable disagreement regarding the significance of phytates in nutrition, and the information on this subject is very confusing. Even though the data were not yet convincing, the British Government, during the recent war, added calcium carbonate to all flours in order to counteract the possible effect of phytates upon the metabolism of calcium in the British dietary. There is reason to believe that this precaution was unnecessary. From time to time, nutrition experts and others have advised against the use of large amounts of cereal products in the dietary on the grounds that the phytates contained in these cereals interfere B-2 (con't.) -2- with calcium metabolism. The poor or the world must eat diets high in cereal content. Thus, it is or considerable importance that we establish the significance of the effect of phytates upon mineral metabolism. We wish first to study the effect phytates on iron metabolism. In the past it has been difficult to measure the effect of a factor upon mineral metabolism because the techniques for measuring mineral absorption and excretion have been too crude. The recent production of radioactive minerals by cyclotron bombardment offers radioactive minerals are metabolized in exactly the same manner as ordinary minerals. Thus, it is possible to tag iron atoms, combine the iron into a compound, feed it and follow its metabolism in the body. Very minute quantities of radioactive substance are required, and the radioactive emanation from this substance is also extremely small. The emanation is of the same physical character as cosmic rays and the quantities which we would give emit less energy than from the cosmic rays continually bombarding people living at the altitude or Denver, Colorado. This applied only to the regions where there are stores of iron in the body. In connection with blood preparations for the armed forces much larger quantities (3 to 5 times) of radioactive substances have been given to more than 100 medical students. Their blood was checked continually over a period of two years. There were no clinical signs attributable to this radioactive material. There is absolutely no ground for caution regarding the quantities of radioactive substances which we would use in our experiments. The fact that Dr. George Whipple has used these compounds in much greater quantities in his clinical investigations at the Rochester Medical School confirms this statement. We had first planned to investigate the effect of phytates upon the metabolism of iron and calcium using rats. This would have been better because large numbers of animals could be used and extensive analyses could be performed on various tissues. Unfortunately, data obtained on rats would be of little interest in human nutrition for two reasons. The rat can utilize ferrous and ferric iron with apparently the same ease, whereas human subjects utilize ferrous iron two to twenty times better than ferric iron. Thus, rats differ from humans in their metabolism of iron. Secondly, there is present B-2 (con't.) -3- in the intestine of rats an enzyme called phytase which has the ability to break down the phytate molecule, whereas the intestinal tract of human beings contains little or none of this enzyme. Thus, rats differ from humans in the manner in which they metabolize phytate. Because we are interested especially in establishing the significance of phatates in human nutrition, it will be necessary for us to use human subjects. Since it has been shown that iron is absorbed by normal subjects most rapidly during the period of active growth, it is advisable that children be used as experimental subjects. Since human beings vary greatly in their metabolism or these substances, it is necessary that the same subjects be used for the entire series of experiments. Since the previous dietary has an influence upon the absorption and metabolism of minerals, it is important that the previous diet of all subjects used in an investigation be the same. Thus, the experiment requires that we use children who have been in the same institution for at least one month. In selecting the children, it is important that they have no disturbance or abnormality which would interfere with the metabolism of foods. During the month or January, we propose to run an animal experiment using radio-iron so that we may work out our techniques not possible today to give you a detailed outline of the clinical procedure which we would follow. It is likely that we will cant to give the subjects a vitamin supplement during the month or January to make certain that they have no malnutrition, for this might interfere with the absorption and metabolism or iron. On or about February 1, we expect to be ready to initiate the clinical experiment. This experiment will be divided into five periods. On the first day of the experiment, we would give to each subject several micrograms of radio-iron in the form of ferrous sulfate in an aqueous solution. This feeding would be given early in the morning to subjects who have fasted overnight. Periodically during that day, we could take samples of blood (no more than 5 ml). During the next two or three days, similar samples would be withdrawn. The radio-iron content of the plasma and red cells of each sample would be determined. In this way we can obtain a record of the rate and amount of iron absorption. B-2 (con't.) -4- After several days have elapsed so that the subjects will be stabilized again, the second period could be started on the same subjects. The second experiment is identical to the first except that a measured quantity of sodium phytates will be fed in the solution containing iron. Blood samples will be withdraw and analyzed as before. This run will demonstrate the effect of sodium phytate upon the absorption of iron. In the third period, the same amount of radio-iron will be administered and the same quantity of phytate will be given. However, the phytate will be fed in the form of rolled oats, corn or whole wheat bread. Blood samples will be withdrawn and analyzed as in the other two periods. This experiment will demonstrate to the effect of phytates in food upon the absorption of iron. In the fourth experiment, the same quantity of radio-iron will be fed and this time a diet consisting of the average American diets will be fed. Blood samples will be withdrawn and analyzed as before. This experiment will demonstrate whether the amount of phytate in the average diet has a significant effect upon the utilization or iron. The fifth experiment will be a repetition of the first in order to establish whether the experimental subjects have changed during the period of the investigation. We believe that these five experiments will take about eight weeks to complete if there are no unforeseen difficulties. In order that we may be certain that ten subjects complete the experiments, we should perhaps start with 15. We are planning this investigation so that it will not interfere with the routine of your institution. If there is any way in which we can reward our subjects, we would be glad to do so. If the experiment can be carried out in your institution, we will be glad for any assistance which your medical staff can give use. Of course, we would recognize this assistance when the results are finally published. We hope that this experiment can be carried out in your institution because the children there would be B-2 (con't.) -5- ideal for this investigation and because you are convenient to the Massachusetts Institute or Technology Laboratories there the analysis of blood samples will be carried out. Professor Robley Evans of our Physics Department is collaborating with us, and the radio-iron will be prepare in the M.I.T. cyclotron. I assure you that the results of this investigation will be of great importance and will influence our thinking in terms of the nutrition of mankind. Sincerely yours, Associate Professor or Nutritional Biochemistry RSH:maa B2/A DRAFT COPY March , 1949 Dr. Paul C. Abersold Isotopes Branch U.S. Atomic Energy Commission P.O. Box E Oak Ridge, Tennessee Dear Paul: This letter is to accompany, and to recommend to you for approval, the application by Dr. , Medical Director of the Walter Fernald School, and Professor Robert S. Harris of M.I.T. for Ca-45 to be used in some important fundamental experiments in nutritional biochemistry on selected young human subjects. All facilities for carrying out the proposed experiment are at hand. The experiments parallel a similar previous study by Prof. Harris and Dr. Sharpe using radioactive iron of cyclotron origin. The present calcium experiments were originally planned Bell over a year ago. Since then an exhaustive study of the many factors involved in this calcium experiment has been made using rats, guinea pigs, and dogs. as the experimental subjects. The overall result of this work has been the perfection of experimental techniques in dealing with Ca-45, the experience gained by the research team which is to handle the human nutritional problem, the verification of a number of physical factors relating to calcium uptake and retention, and some detailed knowledge of the degree of nonuniformity of deposition of radiocalcium in various parts of the skeleton. In this connection, for example, Mr. Robert Dudley of my laboratory and Dr. Brown Dobyns of the Massachusetts General Hospital have recently submitted for publication in Science a paper relating to the techniques which they have developed and some of the results obtained in studies of the nonuniformity of deposition of radio-calcium in the skeleton of dogs. By means of calibrated autoradiographies microphotometered in order to determine local variations of radiation dosage, Dudley and Dobyns have shown that local concentrations of radioactivity can give rise to "hot spots" of dosage, and that the dosage rate in such hot spots is about six to ten times the value which would be obtained if all the radioactivity were uniformly distributed in the bone. I have been personally responsible for at least a year's delay in carrying this experiment on to the case of human nutrition. My feeling was that detailed animal experience should be obtained first. I am now satisfied that this has been done adequately, and that tee proposed human experiment can be carried out without going above trivia dosage levels. B-2/A (con't.) Four simple approaches to the conclusion that the dosages would be adequately small are as follows. 1. It is planned that each human subject will receive 1 uc of Ca-45, mixed with the 250 mg of calcium which is the content of the normal single meal under experimental study. All patients would be on calcium balance and would be receiving the normal 750 mg of calcium per day. From previously available data on humans, obtained by nonradioactive methods, it appears that about 25 percent to the administered calcium may be retained in the growing human. This factor is, if anything, slightly higher than would be expected from our observation of the calcium retention in a young growing dog to which calcium of high specific activity was administered. Assuming, then, that 0.25 uc of calcium is retained in the skeleton. it follows from the so-called "60-formula" that the average bone dose would be expected to be 0.25 millirep/day. This is just equal to the dosage at sea level from cosmic rays and local gamma radiation. It will be remembered that the dosage rate in the blood stream is due to the potassium content of red cells is 0.1 millirep/day. Taking hot spot concentrations as giving as inch as ten times this average dose, the highest local dosages expected would be of the order of 2.5 millirep/day. 2. As Ca-45 has a half period of 160 days, the time integral of the dosage would amount to only 0.6 in the hot spot areas, and only 0.06 rep of average dose to the bone. 3. One might ask approximately what quantity of radium should be swallowed in order to give a radiation dose which is roughly comparable to that given by 1 uc of Ca-45. Recalling that the average beta ray energy of Ca-45 is about 0.08 Mev, and that the total alpha ray energy of Ra, Rn, Ra-A, Ra-C totals about 15 Mev, taking a low relative biological effectiveness of about 5, and making the conservative assumption that the fractional uptake of calcium might be as much as ten times the fractional uptake of radium, one would infer that 1 uc of Ca-45 would be the approximate equivalent of (0.08/15) x (10) x (1/5) = 0.01 uc of Ra. In the field of radium poisoning, where the smaller amounts of fixed radium which have been observed to have deleterious effects are of the order of 1 uc of radium, and where the fractional uptake is usually of the order of 2 percent, the swallowing of 0.01 uc of radium would probably never be viewed with alarm. 4. Karl Morgan's calculation (J. of Phys. and Colloid. Chnem. 31, 984 (1947)) indicated a drinking water tolerance of 3.5 x 10-3 uc/ml water. The average daily water intake being about 1.5 liters, this would amount to a daily intake of about 5 uc. Because the presently proposed nutritional experiment contemplates a single administration of 1 uc, I should be inclined to regard it as perfectly safe. 2 B-2/A (con't.) Because these rough estimates of the dosage all lead to such extremely small values, I have thought it unnecessary for Dr. _________ and Prof. Harris to encumber their application by appending a full report on the animal experiments which have been carried out during the put year. However, I am sure that the animal data can be supplied promptly in case you or other feel that they should be submitted. Cordially yours, Robley D. Evans Professor of Physics 3 B-3 The Commonwealth of Massachusetts Department of Mental Health Room 701, 100 Nashua Street, Boston, 14 December 27, 1945. Malcolm J. Farrell, M. C., Supt. Walter F. Fernald State School Waverley, Massachusetts Dear Dr. Farrell: Acknowledgment is made of receipt of your letter and attachments or December 22nd, concerning a proposed piece of research in connection with the Nutritional Biochemical Laboratories of the Massachusetts Institute of Technology. Perhaps you have not yet become familiar with our procedure in research activities. These proposed activities go through the Advisory Council on Research and Teaching and it well may be that that Committee would invite you to be present at the time when they consider this proposal. Dr. Yakovlev has sat in at those meetings from time to time and undoubtedly would clarify the situation for you. I believe that the next meeting is on the third Thursday of January. Pending the official meeting I see no objections to your selecting the fifteen or so patients, obtaining the necessary family approvals, and going through the preliminary stage of giving them the extra vitamins during January prior to the actual research work to start early in February. I'll send your Protocol to Dr. Tadgell as Secretary of the Advisory Committee and have no doubt but what it will be considered at the next meeting. Very truly yours, Clifton T. Perkins, M. D. Commissioner CTP:HFT B-4 The Commonwealth of Massachusetts Department of Mental Health COMMITTEE ON PSYCHIATRIC EDUCATION AND RESEARCH December 29, 1945 Malcolm J. Farrell, M. D. Walter E. Fernald State School, Waverley, Massachusetts Dear Dr. Farrell: Doctor Perkins has forwarded to me the material concerning the proposed research study to be made in conjunction with the M. I. T. Nutritional Biochemistry Laboratories. This matter will be brought before this Committee at its next regular Meeting which is scheduled for Thursday, January 24, 1946. As Secretary, I am taking the liberty of inviting you to be present. The informal Meeting will be held at the Hotel Kenmore, Boston, at 5:30 P. M., in the lounge-room. Dinner will take place at 6:30 P. M., and, after dinner, the Committee will repair to the Boston Psychopathic Hospital (Dr. Solomon's office) for the Scientific Session. I trust you can arrange to be with us. An official notice will be sent to you about the middle of January. Greetings of the Season. Very truly yours, Sgd. M.D., Secretary HAT:SAG B-5 The Commonwealth of Massachusetts Department of Mental Health Room 701, 100 Nashua Street, Boston, 14 March 29, 1946 Malcolm J. Farrell, M. D., Supt. Walter E. Fernald State School Waverley, Massachusetts Dear Dr. Farrell: On March t, I received a copy of the Minutes of the Meeting of the Advisory Committee on Psychiatric Education and Research, which meeting was held at the Boston Psychopathic Hospital on January 24, 1946. This was the meeting at which you discussed the proposed research work of a collaboration nature between the Walter E. Fernald State School and the Nutritional Biochemistry Laboratories at the Massachusetts Institute of Technology. The proposed research was unanimously approved by the Advisory Committee, and the department concurs in that approval. It is my understanding that you have already encouraged some preliminary steps and I see no reason for your not proceeding in the research at the full and proper level. Sincerely yours, Clifton T. Perkins, M. D. Commissioner CTP:HFT