EXHIBIT 58-LETTER AND REPORT FROM LEBERCO LABORATORIES, ROSELLE PARK, N.J., TO E. FOUGERA & Co., INC., JANUARY 3, 1958, BE ANIMAL STUDIES WITH ORABILEX LEBERCO LABORATORIES, Roselle Park, N.J., January 3, 1958. Submitted to: E. Fougera & Co., Hicksville, Long Island, N.Y. Assay number: 70314 Date received: November 25, 1957. Test material: Orabilex powder; no code number. Ten normal, healthy dogs of mixed breed were used as test animals in this experiment. The anmials were divided into three groups of three test animals and one animal as control. Each group received a different level of the test material. Group one received a level of 100 miligrams per kilogram of body weight. Group two received 1 gram per kilogram of body weight. Group three received 1.5 grams per kilogram of body weight. Upon admittance to our laboratory the dogs were placed in quarantine for a two-week period: During this time they were examined by our staff veterinarian for any diseases or anatomical abnormalities. The animals were all dewormed with Anthol (Jensen-Salisbry). All of the animals received the recommended doses of canine distemper vaccine and infectious canine hepatitis vaccine (Pitman Moore). At the end of this preliminary treatment period all the animals had a normal appetite, presented a healthy appearance and bowel elimination. They were then placed on the experimental regime. All the dogs were housed in individual tile pens having level wooden floors on a slanting slate frame. The urine was washed away continuously through a trough containing running water. The cages were cleaned daily and the animals allowed to exercise in a run for an hour per day. The animals were maintained on a mixture of one part kibbled food and one part fresh cooked ground-horsemeat. Fresh water was available at all times. Each animal received approximately 500 grams of the diet each morning. Any food that was not eaten within one hour was removed. The test material was weighed out in relation to the body weight of each animal. The weighed doses were placed into number twelve gelatin capsules. The filled capsules were placed into each of the respective dogs' mouth beyond the back of the tongue; thus the technician was positive that the entire dose was ingested. The medication was administered daily in the late afternoon for a period of two weeks. A liver function test (Hanger's cephalin flocculation) and a kidney function test (Rowntree and Geragaty; Phenolsulfonephthalein test as described in Kolmer, Approved Laboratory Technique, 5th edition) was performed on each animal initially and again when the experiment was completed. At the end of the experimental period the animals were sacrificed and carefully examined for any morphological changes. Tissue consisting of the thyroid, liver, kidney, spleen and duodenum was carefully removed, weighed and sections placed in Bouin's solution for fixation prior to histological study. MACROSCOPIC EXAMINATION OF ORGANS REMOVED FOR HISTOLOGY 100 mg. per kilogram All three animals in this group had a normal liver both in size and color (chocolate). The surfaces were clear and no adhesions were apparent. The kidneys of each animal were normal in appearance and size. The capsule could be removed easily with no adhesions to the surface occurring. The cut section showed a distinct delineation between the cortical and medullary structures. All of the spleens were normal in size, color, consistency, and a clear surface. The thyroid glands in each dog were somewhat larger than normal and darker in color showing some congestion. Dogs 1 and 2 showed a normal duodenum. The mesentery and adjacent lymph nodes were normal in size and appearance. The cut section showed a normal color and wall thickness. Dog number 3 showed a thickened intestinal wall and some mucosal irritation. The lymph nodes adjacent to the duodenum were slightly enlarged and inflamed. Intestinal worms were found in the lumen which may have caused this irritation. is no basal membrane, the cells rest on the reticular connective tissue. Through- The method employed is that known as Rowntree and Geraghty as described Percent of phenolsulfonephthalein eliminated The feeding of the test material, Orabilex powder, did not seem to affect the general activity of the animals. The animals receiving 1 gram and 1.5 grams per kilogram of body weight showed some loss of weight with one animal in the higher dose level dying. Two animals that received 100 milligrams per kilogram of body weight showed an increase in weight over the experimental period while animal No. 3 showed a decrease. At autopsy macroscopic examination of all the organs showed no damage of any type. Histological examination of the kidney, spleen, duodenum, thyroid gland, and liver also showed no change from the normal. The test material when administered as described above does not appear to cause any damaging effect upon the dogs. LEBERCO LABORATORIES, ANNE WOLVEN, A.B., IRVING LEVENSTEIN, PH. D., Director. Mr. FOUNTAIN. The subcommittee staff advises me that they have found nothing in your files to indicate that FDA is aware of certain inconsistencies in this report which raise serious questions concerning the reliability of these data. At this point I would like to ask Mr. Gray to describe these discrepancies as best he can, because he is thoroughly familiar with them, and has gone through them. After his description, Dr. Vos, I would like to ask you to comment on these inconsistencies, and give your explanation of the FDA's failure to detect them. 3 Mr. Gray. Mr. GRAY. Mr. Chairman, the report referred to was prepared by Leberco Laboratories for E. Fougera & Co., and is dated January 3, 1958. It describes the original toxicity tests on 10 dogs, divided into groups of 3 test animals at different dosage levels, and 1 control animal. It is noteworthy at the outset to point out that the internal organs of the control animal were described in great detail in this report but those of the experimental animals were only described as "normal." The report is divided into two sections. The first covers the macroscopic or gross examination of the organs of these dogs, by the naked eye, and the second covers the microscopic or histological examination of tissues from these same organs. In the description of the macroscopic or gross examination, the following statement is made: Dog No. 10 was dead on the ninth day of the experiment and an autopsy could not be performed because rigor mortis had completely set in. However, in the microscopic examination section of the report there is a tabular presentation describing tissues from the organs of the dogs which lists the kidney, spleen, duodenum, thyroid gland, and liver of dog No. 10 and characterizes each of them as "normal." Tissues from these organs could not have been examined and found normal if the dog was not autopsied as previously stated. Similarly, the macroscopic, or gross examination section of the report says of thyroid glands of the dogs in each group that: The thyroid glands in each dog were somewhat larger than normal and darker in color, showing some congestion. However, in the microscopic section of the report, the thyroid gland of each dog is listed and described as "normal.' It is difficult to understand how they could be found larger than normal, darker in color, and somewhat congested on gross examination, and still be described as normal on microscopic examination. Mr. FOUNTAIN. Those are the discrepancies which were noted by Mr. Gray, and he gave some of his own observations in connection therewith. We would like very much, Dr. Vos, to have you comment on these inconsistencies and to give us your feeling as to why the FDA apparently failed to detect them. Or if they did detect these inconsistencies we would like to hear whatever comments you may have concerning them. Dr. Vos. The first point made, that the microscopic description is given in detail for the control animal, but not in detail for the experimental animals, I believe that is undesirable. We certainly should have had a more detailed description of the microscopic findings on the experimental animals. However, if they were essentially normal, that conveys the information we need. We could evaluate it more critically, certainly, if the details were given. It is not the optimum way of making a report. Mr. GRAY. Would you say that it is unusual, Dr. Vos, that for the control animal, which did not receive the drug, all of the organs were described in great detail, but in the test animals the organs were not described in any detail at all? Is that not rather strange? Dr. Vos. It is unusual. There is a wide range in the detail that different pathologists submit. Mr. GRAY. Normally wouldn't it be either one way or the other, all, including the control, would be described in detail, or none would be described in detail? Is this not normally the case? Dr. Vos. I would say ordinarily there is some consistency that where there is a lot of detail given, it is evenly distributed over the experimentals and controls. But as I say, there is no set formula for reporting pathological data and there is a great variation between the pathologists. One person will give a very brief summary and others will go into extreme minutia. The next question you raise is the fact that for dog No. 10, which died on the 9th day, a report of his tissues, a histological examination, appears. I would agree with you that that is an impossible situation, and if our reviewer saw that, he didn't notice it, or didn't comment on it or criticize it. It may well have escaped his attention. I don't know. My interpretation of it would be carelessness by the reporter in typing it up. Mr. GRAY. What would it have been typed from? If the dog was not autopsied, a description of the organs could not have been written up for typing. Dr. Vos. Well, it is an impossible report. Whether it is carelessness or deliberate Mr. GRAY. In other words, it was not followed up, so you have no knowledge of why this inconsistency is present? Dr. Vos. No, we have no knowledge, that is right. Mr. GRAY. Before we go to the next one, let me follow up one further point. Since dog No. 10 died while under test with the drug, would it not normally be considered even more important that this dog be autopsied, in order to determine the cause of death and any possible connection with the test drug, than for a dog that lived and was later sacrificed to be autopsied! Dr. Vos. As you phrase your question, it would be desirable. But this is a normal procedure. In a dog in which postmortem changes have occurred, there is so much change, degeneration after death of the animal, that it is quite common not to make any histological examination of an animal which has been found dead after a long enough time that rigor mortis has set in. Mr. GRAY. I don't believe there was any indication of how long the dog had been dead. Dr. Vos. Well it says, "An autopsy could not be performed, because rigor mortis had completely set in." Mr. GRAY. Do you agree with the statement that rigor mortis makes it impossible to do an autopsy! Dr. Vos. It wouldn't make it impossible to do a gross autopsy. The organs could be examined grossly, but any findings on histological or microscopic, those are synonymous, any findings of microscopic examination of the organ Mr. GRAY. Would it be fair to summarize your position by saying that you might find less in a dog autopsied after rigor mortis had set in, than if you autopsied him immediately after death! Dr. Vos. No; I am saying it is that almost a universal practice. not to make histological examinations of animals which are found dead as long after death as is evidenced by the fact that rigor mortis had completely set in. Mr. GRAY. How long after death does rigor mortis normally set in! Dr. Vos. In a matter of perhaps 4 hours, something of that sort. Mr. GRAY. Then any time a test animal dies during the night and more than 4 hours elapses before he is found, FDA would not feel it necessary for the company to autopsy that dog to attempt to determine. the cause of death! Dr. Vos. When you say autopsy, I consider that to be a gross examination of the organs. We routinely do a gross examination of animals found dead. But in our own laboratories we with, I would say, very rare exceptions we do not make microscopic examination of the tissues of animals found dead and in which rigor mortis has taken place. If there is one special animal we want to get the most possible. information out of that we can, we may try to make some histological examination. Mr. GRAY. Would you not think that a test animal which dies while under test with a drug fits into the category you just described; i.e., one special animal which you think it might be helpful to get the most possible information on! Dr. Vos. I don't think this would be Mr. GRAY. You are saying you don't think an autopsy was justified in this case, is that right? Dr. Vos. I am saying it is not ordinary practice among pathologists, either outside of the Government or within the Government, to do histological examinations of animals found dead in this manner. There is so little value that can be obtained from the effort that is put into it. In other words, there is so much confusion between the changes of postmortem degeneration, and the possible effects of the drugs, that trying to separate those is almost a thankless task. You can make some guesses, but Mr. GRAY. Would you say that it is not even worth the effort, then! Dr. Vos. That is the general feeling. |