The "Bressler Report"

       Note: This is the text of an FDA report on Searle
                (Part 5 of 5 ... Pages 69 - 76)

EIR 4/25/77 to 8/4/77                      Searle Laboratories
JSA/DME/JT/LF                              Div. G.D. Searle & Co.
                                           4901 Searle Parkway
                                           Skokie, Illinois 60076
(Page 69)

When this study was reviewed by the Bureau of Foods in l975, the
dose-related incidence of uterine polyps was noted.  The appropriate
slides were requested by FDA at that time and were reviewed by three
groups of pathologists:  1.)  The Division of Pathology, Bureau of Foods,
2.) Armed Forces Institute of Pathology, 3.) Massachusetts Institute of
Technology.  Copies of the reports submitted by the 3 groups and related
correspondence were obtained and are attached as exhibits #43-45.

Dr. Rudolph Stejskal was responsible for the microscopic findings and
accuracy on these findings in the submission to FDA.  Only Dr. Stejskal's
name appears on the submission.  However, a Dr. Joseph H. Smith, M.D. also
read slides for this study and his initials appear on some of the
microscopic examination sheets.  Dr. Frith questioned some of the
terminology used in describing tissues.  Dr. Stejskal stated that Dr.
Smith had come directly to SEarle from a hospital situation.  Due to his
human pathology background, his description of animal tissues was somewhat
different than that used by veterinary pathologists.

Dr. Stejskal joined SEarle in July of l973, therefore, he had no input
into the pathology protocol, since E-77/78 was initiated in November of
l971.

No microscopic worksheets or other "raw data" relating to microscopic
pathology could be found for study E-77/78.  We were told by Searle
personnel that the original microscopic findings were dictated by the
pathologists (Stejskal & Smith) onto belts, and then typed onto sheets
which were placed in a binder.  The belts were then discarded and
apparently the bound microscopic pathology sheets were either discarded or
lost, after the study report was written.  Therefore our verification of
the microscopic findings submitted to FDA was limited to a complete
inventory of the slides and tissue blocks and microscopic examination of a
representative number of slides by Dr. Frith.

Our inventory of the slides and tissue blocks for each animal
included a complete list on the tissues sectioned, the number
of slides made from each tissue, and a complete count of the
total number of slides and blocks for each animal.  We also checked
the identification numbers on every slide and tissue block.  We
examined a total of 7,872 slides and 7,360  tissue blocks.  The
average number of organs submitted for tissue processing was

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 20 per animal.  No errors in slide identification were noted,
although in many cases the number of organs submitted for sec-
tioning was less than specified in the protocol.  A detailed dis-
cussion of this can be found under the heading PROTOCOL.

In addition to the discrepancies noted by Dr. Frith, some other errors
were noted in the submission to FDA.  A mammary tumor found in rat F27CF
was described as a papillary cystadenoma on the individual pathology sheet
(page 105, Volume II of the submission to FDA) and as an adenocarcinoma on
the summary table 12, page 96, Volume I of the submission to FDA.

Page 92, Volume I of the submission to FDA (a summary table) reports that
animal J23CM was found dead after 754 days on study, while the individual
pathology sheet for this animal (page 56, Volume II of the submission to
FDA) reported that the animal was found dead after 620 days on study.
The correct figure is 620 days, since J23CM was placed on the study on
11/17/72 and was found dead on 7/29 /73.

In several instances the histopathology technician made notes at the
bottom of the gross pathology sheet to indicate that certain organs were
not present in the bottle of fixative.  (and were therefore not available
for sectioning).  Yet in three of these instances (animals A4CM, K23CF,
and J3CM) a diagnosis appears in the submission to FDA.

CHARTS, DIAGRAMS AND TABLES

It was necessary to construct a number of charts, diagrams and tables to
facilitate our review of the data.  For example we constructed a chart, by
housing group, showing the identification and complete pathology history
of each of the 360 animals.  We also rearranged this chart into dosage
groups, a copy of which is attached as exhibit #35.

To compare survival data it was necessary to construct a survival table.
This also involved devising a calendar to show days and weeks on study for
each housing group, taking into account the starting dates for each group.
This also included tables showing the numbers of days and weeks animals
were on study and a table comparing the survival data from various
sources.

We constructed a chart showing diet calculations (gm./kg) and total
amounts of DKP used (gm./batch).  This is attached as exhibit #30.

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Three tables were constructed which summarize the FDA statistical analysis
of body/feeder weight data.  They are attached as exhibits #39-41.

All of the charts, diagrams and tables that we constructed are attached to
the report as exhibits and are referred to in various sections of the
report.

EXHIBITS

#1.  G.D. Searle & Company Annual Report for l976.
#2.  Organizational Chart of Pharmaceutical/Consumer Products Group.
#3.  Organizational Chart of World Wide Pharmaceutical R&D Group.
#4.  Organizational Chart of Preclinical R&D Group.
#5.  Organizational Chart of Product Safety Assessment Group.
#6.  Copies of Computer-Generated Randomization Tables used by Searle
     to assign the Dose & Housing Groups.
#7.  Diagram showing Typical Housing Group of 30 animals, containing
     a random distribution of control and treated animals.
#8.  Diagram showing arrangement of food cups on cart, used in feeding
     the animals.
#9.  Copy of "Glossary of Terms for Aspartame and its Diketopiperazine"
     and "Analytical Data and Specifications of Food Grade Aspartame".
#10. Copy of shipping labels for rats received from

#11. Copy of protocol with amendments for Study P.T. 988S73 (E-77/78).
#12. Copies of CV's for principal persons involved in study E-77/78.
#13. Copies of Batch Records for the manufacture of DKP, lots 1R through
     5R.
#14. Copies of pages from SEarle chemist Jack Drogt's notebook, con-
     cerning the manufacture of DKP.
#15. Copies of Analytical Reports for DKP, lots 1R through 7R.
#16. Copy of Searle memorandum dated 12/4/69, concerning DKP Specifica-
     tions.
#17. Copy of DKP Specification Sheet (not dated) entitled
     "Tenative Specifications for SC-19192".

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#18. Copy of DKP Specification Sheet entitled "Specifications for
     SC-19192, Specification #C40606C".
#19. Copies of pages 75-84 & 285 of lab notebook #AR-39, concerning
     assay of DKP, lots 1R, 2R & 3R.
#20. Copies of pages 60-63 of lab notebook VSH-I, and page 269 of
     lab notebook book #AR-23, pertaining to analysis of DKP lot 4R.
#21. Copies of pages 250, 251 and 257 of lab notebook #AR-57, and
     pages 44-49 of lab notebook #AR-68, pertaining to analysis of
     DKP lot 5R.
#22. Copies of pages 83-86 of lab notebook #AR-77, concerning analysis
     of DKP lot 6R.
#23. Copy of page 31 of lab notebook #AR-93, concerning analysis of
     DKP lot 7R.
#24. Copy of protocol for DKP stability study, dated 1/13/72.
#25. Copies of pages 51-56 of laboratory notebook #AR-49, assigned to
     C. Seul.  These pages describe a preliminary TLC Test for
     recovery of DKP from the diet mixture.
#26. Copies of pages 53-59, 67-72, 88-89, 106-107, 144-145, 156-157,
     and 284-285 of laboratory notebook #AR-51, assigned to Barbara
     Bickford.  These pages refer to the assay procedure and methods
     for the DKP Stability Study.
#27. Copies of Analytical Reports for DKP Stability Study.
#28. Copies of DKP Compound Inventory Cards.
#29. Two photographs showing a non-homogeneous sample of DKP diet mixture.
#30. Chart showing diet calculations (gm.kg.) and total amounts of DKP
     used (gm./batch).
#31. Two memos dated 7/14/77 from Thomas F.X. Collins concerning interview
     with Ray Schroeder.
#32. Memo dated 7/19/77 from Thomas F. X. Collins describing the 7/18/77
     interview with Ray Schroeder.
#33. Memo of Telephone Conversation between Jerome Bressler and Attorney
     John H. Bickley Jr., dated July 25, l977.
#34. Copies of records concerning calculation of diet concentrations, food
     concentrations prediction records, dates of bath mixing, and calcu-
     lation of mean food intake values.
#35. Charts organized by dose group, showing the identification and
     pathology history of each of the 360 animals on study.
#36. Memo dated April 5, l976, from Dr. John H. Rust to Dr. R. McConnell.
#37. Searle memo dated September 30, l974, by Dr. McConnell.

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#38. Memo dated August 29, l974 from Dr. G. L. Schoenhard to Dr. K.S.
     Rao.
#39. Table 1 - Summary of Average Body Weights and Weight gain (%change/
     week) from the FDA Statistical Analysis.
#40. Table 2 - Summary of food intake (g/day and g/kg./day and dosage
     (mg./kg./day) from the FDA STatistical Analysis
#41. Table 3 - Summary of Food Efficiency (g. gained/100g. actual food
     consumed) calculated in the FDA STatistical Analysis.
#42. Computer printout of FDA Statistical Analysis of food intake and
     body weight data.
#43. Pathology report from Division of Pathology, Bureau of Foods, con-
     cerning uterine polyps, along with correspondence, and memo from
     Janet Springer.
#44. Pathology report from Armed Forces Institute of Pathology, concerning
     uterine polyps.
#45. Pathology report from Massachusetts Institute of Technology, concern-
     ing uterine polyps.
#46. Written account of interviews with Dr. Jean Taylor on 6/2/77, 6/3/77,
     and 6/7/77.
#47. Written account of interview with Judy Beauchamp on 6/2/77.
#48. Written account of interview with Barbara Bickford on 6/1/77.
#49. Written account of interview with Clifford J. Seul on 6/2/77.
#50. Written account of interview with Bartolome R. Tangonan on 6/1/77.
#51. Written account of interviews with Tony Martinez on 5/19/77,6/3/77,
     7/7/77, and 8/2/77.
#52. Written account of interview with Ted Reichert on 5/24/77.
#53. Written account of interview with Barbara Bickford and Clif Seul
     on 6/2/77.
#54. Written account of interview with Judith Schmal on 6/2/77 and
     6/7/77.
#55. List of animals bled at 104 and 114 weeks.
#56. Written account of interview with Alan Mitchell on 7/20/77.
#57. Written account of interview with Raymond G. Schroeder on 7/18/77.
#58. Injection records showing administration of penicillin, dates of
     administration, rat numbers, and units injected.
#59. Methodology for "Phenistix" determination of phenylketones in urine.

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#60. Dr. Frith's report of examination of slides for DKP study
     (E-77/78).
#61. Key for animal identification card numbers used on the body/feeder
     weight teletype sheets.
#62. Chart correlating animal cage numbers with pathology numbers,
     arranged by dose group.
#63. Chronological list of pathology numbers and corresponding animal
     cage numbers.
#64. Organizational charts showing responsibility during the time that
     study E-77/78 was conducted.
#65. Volume entitled "tissue masses & deaths".  Chronological list
     of all animals that died, during study, and dates that masses were
     first observed.
#66. Charts of days on study for each animal.
#67. Calendar for duration of study showing starting dates, days and weeks
     for each group.
#68. Survival table.
#69. Charts of Housing/Dosage Groups.
#70. "Observations for Drug Effects" records for housing groups A through
     F.
#71. "Observations for Drug Effects" records for housing groups G through
     M.
#72. Ophthalmoscopic records and copies of pathology sheets that have
     ophthalmoscopic findings.
#73. Life table analysis and statistics on body weight and food
     consumption data by Dennis Wilson, Div. of Mathematics, Bureau of
     Foods.
#74. Evaluation of feeding study on DKP, a conversion product of
     Aspartame, by Janet Springer/Ann Ducca, FDA Division on Mathematics.
#75. Volume A - teletype sheets for body and feeder weight data, housing
                groups.
     Volume B -           "
     Volume C -           "
#76. Copy of Searle Computer Program.
#77. Volume of protocols, organ weights, dosage, hematology, urinalysis,
     blood chemistry, and protein electrophoresis.
#78. Complete gross pathology sheets, males.
#79. Complete gross pathology sheets, females.
#80. Key to slide tissue identification numbers and abbreviations.
#81. Key to stain abbreviations.

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#82. Copies of submission appendix tables relating to hematology,
     clinical chemistry, urinalysis, and electrophoresis, along with
     check marks showing errors, and attached copies of raw data
     sheets documenting the errors.
#83. Copies of submission appendix tables for organ weights, with
     errors indicated, and copies of pathology sheets documenting
     the errors.
#84. Data sheets showing the phenylketones test erroneously labeled
     "phenylalanine".

                         Signed by:

                         John S. Arnold
                         Investigator

                         David M. Eerspamer
                         Investigator

                         Dr. Jean Taylor
                         Toxicologist

                         Dr. Leonard Friedman
                         Biochemist

                           (75)

Addendum:

Exhibit #85   - Copy of Volume 1 of the submission to FDA.

Exhibit #86   - Copy of Volume 2 of the submission to FDA, consisting
                of the individual pathology summaries, both gross and
                microscopic.

Exhibit #87   - Statistical analysis of Blood and Clinical Chemistry
                Data by Dennis Wilson, Division of Mathematics, HFF-
                110.

Exhibit #88   - Copies of pages from Histology accession book #770C,
                and Histology inventory sheets.

Exhibit #89   - Documents from Searle's Math-Stat. Dept. concerning
                statistical analysis of study 988S73.


                                  Signed

                                  Jerome Bressler
                                  Team Leader

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