(12)
PROTOCOL AND AMENDMENTS
A copy of the protocol
for this study was obtained and is attached
to this
report (See Exhibit #11).
The protocol includes 4 amendments which are
dated Aug 20, l973, (amendments
#1 and 2), Sept. 6, l973 and Jan 9, l974.
Amendment #1 dated Aug
20, l973 specified 4 additional clinical chemistry
laboratory measurements:
1.) serum insulin, 2.)serum ornithine carbamyl
transferase, 3.) serum
protein electrophoresis, 4.) serum total protein.
Two of the above assays
(serum insulin, and serum ornithine carbamyl
transferase) were apparently
not done, because no data for these two
parameters was submitted
to FDA, and we could find no raw data or other
evidence that they were
done.
Amendment #2 dated Aug
20, l973, specified 8 coronal sections of
brain to
be examined microscopically,
and also described the procedure for
sectioning the urinary
bladder. Four transverse sections from each
urinary bladder were
to be examined microscopically.
Amendment #3 dated Sept.
6, l973 extended the study until it reached
a
point where mortality
reduced the control group to 20 animals per
sex,
provided survival of
treated groups was not less than 10 per sex
per
group. (This represented
a survival of approximately 30%).
Amendment #4 dated Jan
9, l974 added serum cholesterol to the clinical
chemistry measurements
to be made at terminal sacrifice, and terminated
the study after 114 weeks
of treatment. Terminal sacrifice was to begin
on 1-24-74 and continue
through 2-1-74.
Our examination of the
original data showed that serum cholesterol
determinations were done
at day 796 and 798 (terminal bleeding) as
specified in the above
amendment, but the data was not included in
the
submission to FDA. The
submission to FDA (Vol. 1 p. 286) reported
a
significant decrease
in serum cholesterol that was more perceptible
towards the end of the
study, and may have been related to compound
administration. Therefore,
the omitted data may have been important.
Serum cholesterol determinations
were also done at day 546 (78 weeks) and
not reported in the submission
to FDA.
(13)
The protocol for Clinical
Chemistry procedures specified that BUN
determinations were to
be done at 78 weeks (546 days). The submission
to
FDA contained no BUN
data for day 546, but our review of the raw
data
indicated that BUN's
had been done at day 546. Some BUN's were
also done
at day 735 (105 weeks)
and not reported in the submission to FDA,
but this
data was not complete
for all animals.
Attached to the protocol
is a memo dated Oct. 31, l972 which describes
an
acute infection spreading
in the rat colony, and the administration
of
penicillin to combat
the infection, and a memo dated May 8, l973
listing
scheduled dates to be
added to Body and Feeder Weights of housing
groups A
& B.
The final Histology Lab
Protocol, dated 1-21-74, specifies 24 organs
to be
embedded for control
and high dose animals, and 19 organs to be
embedded
for low and mid dose
groups. The organs which were to be embedded
for the
control and high dose
groups but to be omitted in the low and mid
dose
groups include: lymph
node, nerve, bone, eye, and salivary glands.
Pathology sheets (blank
forms) to be used at terminal sacrifice were
reproduced (xeroxed)
with check marks, time (death to tissue fix),
fixative, study, and
project number already entered. Twenty-seven
(27)
organs were checked off,
to be embedded. However, as stated above,
the
control and high dose
animals were to have 24 organs embedded, according
to the protocol, and
the mid and low dose 19. Therefore, all pathology
sheets for animals killed
by design have incorrectly identified the
specific organs and tissues
to be embedded.
In addition to the above
error, in many cases the actual number of
tissues
embedded was less than
the 24 (control and high dose) or 19 (low
and mid
dose) specified in the
final Histology Lab Protocol dated 1-21-74.
Specific figures for
numbers of tissues embedded at terminal sacrifice
are
as follows:
NUMBER SPECI- NO. OF
ANIMALS
ACTUAL ACTUAL FIED IN
PRO- NOT IN ACCORD
RANGE AVERAGE TOCOL WITH
PROTOCOL
CONTROLS 10-24 20 24
129 of 144
LOW DOSE 12-23 19 19
19 of 72
MID DOSE 4-24 18 19 28
of 72
HIGH DOSE 9-25 22 24
51 of 72
(14)
PERSONNEL AND RESPONSIBILITY
The names of Dr. K.S.
Rao, Dr. R. Stejskal, and Dr. R.G. McConnell
appear on the final study
report, indicating that they are the
authors of this report,
and were responsible for the study.
Following are the principal
person involved with study E-77/78 and
their specific areas
of responsibility:
1.) Dr. Robert G. McConnell
- Director, Pathology-Toxicology
Laboratory 1970 through
1974. Dr. McConnell functioned as the Path-
Tox advisor on study
E-77/78. He is no longer employed by Searle.
2.) Dr. Suryanarayana
K. Rao - Manager, General Toxicology
Laboratory, June 1971
until he left Searle in May of 1977.
Dr. Rao was the Path-Tox
monitor for study E-77/78. In
1971 Dr. Rao monitored
30 studies, in 1972 forty-seven
(47) studies, in 1973
twenty-nine (29) studies and in
1974twenty-five (25)
studies.
3.) Dr. Rudolf Stejskal
- Senior Research Investigator,
Pathologist. Dr. Stejskal
was responsible for the micro-
scopic findings and accuracy
of these findings in the study
report of E-77/78. Because
Dr. Stejskal joined Searle in
July, 1973, he had no
input into the pathology protocol.
Also, he did not examine
all of the slides for this study,
but was assisted in that
task by Dr. Joseph H. Smith M. D..
4.) Dr. Joseph H. Smith,
M.D. - Group Leader and Senior Patholo-
gist at Michael Reese
Hospital, Chicago, IL., before joining
Searle in June of 1973.
Dr. Smith examined some of the
slides for study E-77/78,
and supervised the necropsy labora-
tory.
5.) Tony Martinez - Toxicology
Laboratory Supervisor, 1970
through 1973. Mr. Martinez
participated in twelve (12)
studies in 1971, Seventeen
(17) studies in 1972, and
thirteen (13) studies
in 1973. Mr. Martinez supervised
the technicians who worked
on the study. He also
performed some necropsies.
(15)
6.) David K.T. Kie, B.S.,
Research Assistant in Pathology
Laboratory. He performed
some of the necropsies on E-77/78.
7.) Robert Spaet - Research
Assistant. He also performed
necropsies.
8.) Bartolome R. Tangonan
- Research Technician II - He was
involved with preparation
of diet mixtures, daily observa-
tions, weighing and feeding
animals, etc.
9.) Donna K. Helms -
Manager, Safety Evaluation, Project Sche-
duling, Reporting, and
Data Storage, Path-Tox Dept. and
Administrative Assistant
to Dr. McConnell.
10.) Patrica Erdenberger
- Research Assistant, and Histology
lab Supervisor.
11.) Dr. Eugene Joseph
Youkilis - Senior Research Investigator.
He performed the opthalmoscopic
examinations in study
E-77/78.
12.) Judy A Henderson
- August 1972 to present, Research Tech-
nician III, Histopathology
Dept. She was involved with
tissue processing on
study E-77/78.
13.) Judith R. Schmal
- Nov. 1971 to present, Supervisor, Clini-
cal Chemistry Section
of Bioanalytical Laboratory.
14.) Judith A. Beauchamp
- Employed Aug, 1970 to present;
Supervisor Hematology
laboratory since April 1973.
15.) Barbara (Ross) Bickford
- Research Technician, Quality Control
Department. She performed
analyses of DKP diet mixtures for
study E-77/78.
16.) Clifford J. Seul
- Supervisor, Method Development, Stability
Evaluation Laboratory.
He was Barbara Bickford's supervisor
at the time the DKP stability
study was performed.
17.) Jack Drogt - 1967
to present, Senior Research Assistant,
Chemical Development.
Mr. Drogt manufactured the 7
lots of DKP used in the
study E-77/78.
18.) Dr. John E. Dutt
- Math-Stat. Dept.
19.) John Mellman, Math-Stat
Dept.
