Damage to Kidneys, etc. from Methanol
[wood alcohol]:
Renal changes [Table - 3] such as varying degrees of renal tubular degeneration (severe cloudy or vacuolar change) with patchy necrosis were seen in all the cases. Severe congestion of peritubular capillaries (71.42%), moderate glomerular capillary dilatation and congestion (78.57%), mild endothelial swelling (35.7%) and mild mesangial proliferation (35.7%) was the other significant renal findings.
Approximately 3-10% of administered dose of methanol is excreted unchanged by the kidneys[2]. Highest concentration of formaldehyde has been found in the kidney, liver and gastro-intestinal tract by studies using C[14] labelled methanol. Considering that both methanol and its toxic metabolites are concentrated in the kidneys it is not difficult to explain the severe degenerative changes in the renal tubules and peritubular capillaries. Glomerular changes too rescmble the early changes seen in cases of viper bites, Haemolytic uracmic syndrome and toxacmia of pregnancy, all of which have a known toxic aetiology.
Thus changes in the various organs were probably of toxic aetiology, either mediated through the direct toxic action of the metabolite formaldehyde or the metabolic acidosis due to formic acid production.
:: References
1. Bade LK, Snore DP. Methyl alcohol poisoning, Medical News, Medicine and Law as quoted by Ravichandran et a1[8]. 1981; 12:106-108.
2. Barriers GR. Combustion of C[14] labeled methanol in intact rat and its isolated tissues. Amer J Physiol 1950; 163:614-618.
3. Bonnets IL Jr, Cony FH, Mitchell GL Jr, Cooper MN. Acute methyl alcohol poisoning: a review based on experiences in an outbreak of 323 cases. Medicine 1953; 32:431-463.
4. Gender A, Manly H, Churchill D, Hollomby D. Hemodialysis for methanol intoxication. Amer J Med 1978; 64:749-758.
5. Lurid A. Excretion of methanol and formic acid in man after methanol consumption. Quoted by Gender et al[4]. Acta Phamacol Toxicol 1948; 4:205.
6. Kaplan K. Methyl alcohol poisoning. Amer J Med Sci 1962; 244:170-174.
7. Orthner H. Die Methylalkohol vergiftung, as quoted by Gender er al[4]. Berlin: Springer Verlag; 1950.
8. Ravichandran R, Dudani RA, Alongside AE, Chamber KP, Acharya VN. Methyl alcohol poisoning (experience of an outbreak in Bombay). J Postgrad Med 1974; 30:69-74.
9. Zimmerman HJ, Ishak KG. In: Pathology of the Liver." MacSween RNM, Anthony PP, Pscheuer PJ, editors. 1st ed., Edinburgh: Churchill Livingstone; 1979, pp 337.
Below from FDA's Jerome Bressler Report:
ORGAN WEIGHTS
Organ Weights were entered on the gross pathology sheets at the time of
autopsy. We compared all of the individual organ weights on appendix
table 5 in the submission to FDA (Vol 1, pp. 222-226) with the original
data on the gross pathology sheets. A total of eleven (11) errors were
noted in transcribing the raw data from the pathology sheets, to the
tables in the submission to FDA.
The errors are tabulated below:
Wt. Shown In Wt. Recorded on
Animal No. Organ Submission Original Pathology Sheet
A12CM Kidneys 3.75 G 3.45 G
L28LM Ven. Prostrate 747 mg. 474.7 mg.
C0lMM Kidneys 9.40 G 9.219 G.
C02HM Kidneys 1.46 G 4.259 G
E14HM Kidneys 11.74 G 4.746 G
J12HM Pituitary 3.0 mg. 3.3 mg.
J30HM Ven. Prostrate 444 mg. 444.8 mg.
F17CF Ovaries 36.7 mg. 233.5 & 36.7 mg.
H30CF Liver 9.4 G 9.493 G
B20HF Uterus 1115 mg. 1155 mg.
K11HF Adrenals 799.1 mg. 797.1 mg.
_________________________________________________________________
NO evidence was obtained that any attempts were made to determine
whether or not DKP could interfere with any of the clinical lab-
oratory tests conducted. For that matter no information was made
available to us as to whether DKP itself or related compounds did
appear in the blood or the urine of rats fed diet containing this
compound.
teletype sheets referred to previously. In reference to the
low BUN values, Page 29 of the submission contains the following
statement: "BUN" values for the control males at treatment
day 189 were unusually low and may possibly be related to
a technical artefact; as a result, the group mean values for
all treated males at this interval were significantly higher
but, in fact, these values were in the normal range. BUN values
both in control and all treated male groups at treatment
day 364 were unusually low; this again reflects a possible
technical artefact."
(55)
GROSS PATHOLOGY
The pathologists responsible for the microscopic examination (Rudolph
Stejskal and Joseph smith) did not perform the necropsies. Necropsies
were performed by Tony Martinez, David Kie and Robert Spaet, with the two
pathologists avilable for consultation.
The submission to FDA (Vol 1, p. 7) reported that "Rats found dead during
the study were autopsied immediately whenever possible. In cases where
the ncropsy could not be performed promptly, the thoracic and abdominal
cavities of dead rats were opened and the entire animal was immersed in
neutral buffered formalin fixative for subsequent gross examination and
dissection".
Our examination of gross pathology records showed that 98 of the 196
animals that died during the study were fixed in toto and autopsied at
some later date, in some cases more than one year later.
A total of 20 animals were excluded from the study due to excessive
autolysis. Of these, 17 had been fixed in toto and autopsied at a later
date. Following are the twenty animals excluded from the study:
Animal No Date Found Dead Date Autopsied
C21CM 7/3/73 1/11/74
G16CM 9/21/73 1/11/74
G18CM 8/11/73 10/4/73
G26CM 4/2/73 1/11/74
J2CM 5/21/73 1/11/74
J5CM 10/30/72 11/8/72
L10CM 3/29/73 1/11/74
L15/CM 9/9/73 1/11/74
L21CM 4/13/73 1/11/74
L11LM 5/6/73 1/9/74
A14MM 5/21/73 1/9/74
G28MM 1/5/74 1/7/74
J25MM 5/24/73 5/24/73
A3HN 6/17/73 1/9/74
C15HM 1/7/74 1/7/74
(56)
Animal No Date Found Dead Date Autopsied
G13HM 7/25/73 **1/9/74
H24CF 4/29/73 1/11/74
D4HF 7/11/73 7/11/73
D16HF *4/2/73 1/8/74
F6HF 1/5/74 1/7/74
*Although the date found dead was listed as 4/12/73 on the gross pathology
sheet, the "Tissue Masses & Deaths" book listed this date as 4/1/73.
**Although the date found dead was listed as 1/9/74 on the gross pathology
sheet, the "Tissue Masses & Deaths" book listed this date as 7/25/73.
The gross pathology sheet for one of the above animals, F6HF, described a
tissue mass measuring 5.0 X 4.5X2.5 cm. This tissue mass was first
observed on 8/24/73 according to the pathology sheet (Exhibit #79), the
observation records (Exhibit #70), and the palpation record in the "Tissue
Masses and Deaths" book (Exhibit #65). The submission to FDA (Exhibit #8)
reported no tissue mass and the animal was excluded from the study due to
marked autolysis.
In addition to the above twenty animals that were excluded from the study,
many other animals exhibited marked autolysis. For example, D27LF, M25CF,
and H12CF are all described grossly in the submission to FDA as follows;
"all organs examined grossly were markedly autolyzed".
Records for approximately 30 animals showed substantial differences
between gross observations on pathology sheets, when compared with the
individual pathology summaries submitted to FDA. Following is a detailed
comparison of ten of these. (Copies of all the gross pathology sheets,
and the pathology summaries submitted to FDA are attached as Exhibits #78,
#79, and #86).
A2CM
Submission to FDA:
Lung - Focal adhesion
Adrenal - Moderately enlarged
(57)
All other organs examined grossly were unremarkable.
Original Pathology Sheet:
Pituitary - Missing
Lung - Left, mid-portion adheres to the medial area of
the rib cage by a "fibrous" type of tissue. (Sub-
mitted together with relevant portion of the rib cage).
Right, post-caval lobe has undergone consolidation. Contains
grayish-yellowish nodules measuring 2 x 2 mm. (Entire lung
submitted in toto)
Lymph NOdes, Pancreatic - Slightly enlarged
Adrenal - Left, moderately enlarged. Right and left, covered
with tiny yellow spots measuring 1.0 x 1.0 mm.
Lymph Nodes, Mesenteric - moderately enlarged.
Mass - previously described on 8/20/73 has since then regressed.
Prostate - Marked atrophy, all lobes
Seminal Vesicles - Marked atrophy, bilaterally
All other organs examined were grossly normal and unremarkable.
M15CF
Submission:
Mammary gland - subcutaneous mass located in mid-thoracic
region measuring 7 x 6 x 2.5 cm.
Urinary bladder - papillary growth in the lumen.
All other organs examined grossly were unremarkable.
Original:
Mass #1 - Previously described in the left inguinal region
on 2/9/73 has since then regressed.
(58)
Masses #2 and # - Located in the mid-axillary-cervical regions
are all on mass now measuring 7.0 x 6.0 x 2.5 cm
and may be described as irregular in shape, multi-
nodular, smooth-surfaced, non-glistening,
No Spinal Cord yellowish-purpulish in color, non-adherent to the
VL underlying muscle and containing a whitish-yellowish
firm tissue within. (Submitted in toto together with
remainder of tissue).
Heart - Left Ventricle - dilitation and walls thin.
Spleen - Slightly enlarged
Liver - Prominent lobular architecture.
Adrenal - Left, slightly enlarged. Right, unremarkable.
Ovary - Right, small cyst measuring 4.0 x 4.0 mm and
distended with a clear yellow fluid.
All other organs examined were grossly normal and unremarkable.
G10LM
Submission:
Testis - Marked atrophy, unilaterally.
Kidney - Moderate enlargement, mottled appearance, bilaterally.
Small and large intestine exhibited moderate autolysis, no sections
submitted.
All other organs examined were grossly normal and unremarkable.
Original:
Mass which was initially palpated on 2/9/72 (86 days Rx)
in the left inguinal area was actually the left testis which
ascended and went thru weakened left inguinal ring into the
subcutaneous area.
Testis - Left (ascended) appears atrophied (submitted in toto).
Kidney - Moderate, diffuse and uniform enlargement, mottled,
bilaterally (submitted in toto).
Small and large intestines are moderately autolyzed (no sections
submitted).
Thyroid - Moderately enlarged, bilaterally. A 2 mm in dia., dis-
crete, sl raised, moderately firm yellowish-grey lesion
is located in the posterior tip, bilaterally. (Thyroid
submitted in toto wrapped in a lens paper).
All other organs examined were grossly normal and unremarkable.
(59)
L11LM
Submission:
Kidney - Mottled appearance
Testes - Marked atrophy, bilaterally
Prostate - Marked atrophy
All other organs examined grossly exhibited marked autolysis.
Original:
Adrenal - Pale yellow, bilaterally
Kidney - Pale yellow, bilaterally, rough-surfaced, bilaterally,
moderately autolyzed, bilaterally, tiny spaces in the
cortex region measuring about 1 mm in diameter,
bilaterally.
Testes - Marked atrophy, bilaterally, marked autolysis,
bilaterally.
Prostate - Marked atrophy, all lobes
Seminal Vesicles - Marked atrophy, bilaterally
Spleen - Marked autolysis
Pancreas - Marked autolysis
Stomach - Marked autolysis. A glandular portion - numerous, tiny,
pitted ulcerations measuring 1 -4 mm in diameter.
Lymph Nodes, Mesenteric - Marked autolysis
Heart - Wall of left ventricle thin
Brain - Marked autolysis
Pituitary - Marked autolysis
Liver - Marked autolysis
All other organs examined were grossly normal and unremarkable.
M17LF
Submission:
Pituitary - Marked enlargement.
Adrenal - Markedly enlarged and hyperemic, bilaterally.
Mammary Gland - Mass 1, located subcutaneously in left axillary
region, measuring 3 X 3 X 2.5 cm; mass 2, located
subcutaneously adjacent to mass 1, measuring 3 X 2
X 1 cm; mass 3, located subcutaneously in the right
axillary region, measuring 2.5 X 2 X 1 cm; mass 4,
(60)
located subcutaneously in the left inguinal region,
measuring 3 X 1 X 1 cm; mass 5, located subcutane-
ously in the right inguinal region, measuring 2 X
1.5 X 1 cm.
All other organs examined grossly were unremarkable.
Original
Pit - appears markedly hyperemic
Adrenal - Exhibits numerous minute greyish spots on the serosal
surface bilaterally. It appears markedly enlarged.
Mass (1) - A 3 X 3 X 2.5 cm. spheroidal, multinodular, yellowish
white, slightly firm mass located subcutaneously in
the left axillary area. Mass non-adherent to the
surrounding muscles or tissue (submitted in toto).
Mass (2) - A 2.5 X 2 X 1 cm spheroidal, smooth, yellowish white
firm mass located subcutaneously and adjacent to the
above described mass (submitted in toto) mass non-
adherent to the surrounding muscles or tissues.
Mass (3) - A 2.3 X 2 X 1 cm. irregularly shaped, multinodular,
yellowish white, firm mass located subcutaneously on
the rt. axillary area. Mass non-adherent to the
surrounding muscles or tissues (submitted in toto).
Mass (4) - A 3 X 1 X 1 cm. elongated, multinodular, yellowish
white, firm mass located subcutaneously on the left
inguinal area. Mass non-adherent to the surrounding
muscles or tissues (submitted in toto).
Mass (5) - A 2 X 1.5 X 1 cm. flat, multinodular, yellowish white,
firm mass located subcutaneously on the rt. inguinal
area. Mass non-adherent to the surrounding muscles
or tissues (submitted in toto).
All other organs examined were grossly normal and unremarkable.
C1MM
Submission
Kidney Marked enlargement with yellowish discoloration.
Testis Marked atrophy, bilaterally.
(61)
Tissue mass located subcutaneously in the right inguinal area measuring
2.5 X 1 cm.
All other organs examined grossly were unremarkable.
Original:
Mass - Previously described on 12/9/72 and located subcutaneously
in the right inguinal area now measures 2.5 X 2.0 X 1.0
cm and may be described as smooth-surfaced, purplish-
yellowish in color, non-glistening, firm, multi-nodular,
non-adherent to the underlying muscles and containing a
firm yellowish-whitish tissue. (Submitted in toto
together with a portion of the skin and underlying muscle
with remainder of tissue).
Heart - Left ventricle has undergone a moderate amount of dili-
tation. Wall, left ventricle is thin.
Liver - Prominent lobular architecture.
Lung - Right, post-caval lobe-consolidation.
Kidney - Markedly enlarged, yellow and rough-surfaced, bilaterally.
Dilitation of the pelvis.
Adrenal - Covered with tiny yellow spots measuring 1 mm in diameter,
bilaterally.
Testes - Marked atrophy, bilaterally.
All other organs examined were grossly normal and unremarkable.
Tiss. Trimming - Nodules discovered immediately posterior (2.0 cm) to
the pyloric portion of the stomach within the adipose
tissue. Nodules may be described as firm, yellowish
brownish in color. Non-glistening measuring 1.2 X
1.0 mm to 4.0 X 4.0 mm.
E27MM
Submission:
Lung - Moderate diffuse hyperemia.
Eye - Opaque cornea, bilaterally.
All other organs examined grossly were unremarkable
Original:
Lungs - All lobes exhibit moderate diffuse and uniform hyperemia.
(62)
Kidney - Moderate autolysis.
Eye - The entire cornea is opaque, bilaterally.
Spleen - Moderately autolyzed.
Stomach - Numerous 1-2 mm. hemorrhagic ulcerations are located on
the glandular mucosa. Entire small and large intestines
are moderately autolyzed.
Brain & Pituitary - Moderately autolyzed.
All other organs examined were grossly normal and unremarkable.
A1HM
Submission:
All organs examined were grossly unremarkable.
Original:
Testes - Markedly atrophy, bilaterally
Lung, Rt - Middle lobe exhibits a 1 X 1 cm consolidation on the
posterior portion.
Liver - All lobes appear olive green otherwise unremarkable.
All other organs examined were grossly normal and unremarkable.
L27HM:
Submission:
Testes - Right, slightly enlarged; left, mild atrophy.
All other organs examined grossly were unremarkable.
Original:
Testes - lt./appears markedly atrophy
rt./appears to be distended with yellowish white
substance
Seminal V- Appears markedly atrophy bilaterally.
Intestinal - Large, markedly distended with "gas".
All other organs examined were grossly normal and unremarkable.
P.M. Testes - Also, small black areas are noted within along with
the yellowish areas. Black areas measuring 1.0 X 1.0 to
4.0 X 4.0 mm in diameter.
(63)
J30HM
Submission:
Lung - Moderate consolidation of all right lobes.
Testis - Moderate atrophy
All other organs examined grossly were unremarkable.
Original:
Pituitary - Markedly enlarged; slightly hyperemic.
Heart - Left Ventricle has undergone dilitation walls thin.
Lung - Right, anterior, medial and post-caval lobe have
undergone consolidation.
Testes - Marked atrophy, bilaterally.
Seminal Vesicle - Marked atrophy, bilaterally.
All other organs examined were grossly normal and unremarkable.
Dr. Stejskal told us that the other pathologist (Dr. Joseph Smith) who
made microscopic evaluations of the slides, came from a hospital
background (human pathology) and therefore his descriptions and
terminology were a little bit different than one would expect from a
veterinary pathologist.
MICROSCOPIC PATHOLOGY
We have assisted in our review of the Microscopic Pathology of Study
E-77/78 by Charles H. Frith, D.V.M., Ph.D, Director, Pathology Services,
NCTR. Dr. Frith arrived on 6/22/77 and spent 3 days with the FDA team.
He examined slides for a representative number of animals, the selection
of which was made jointly by Dr. Frith and the other members of the FDA
team. A Searle Pathologist was not present during Dr. Frith's review of
the slides. However, Dr. Frith did meet with Dr. Rudolf Stejskal, SEarle
Pathologist, at the conclusion of this review and discussed some of his
findings with him.
The first phase of Dr. Frith's review consisted of the examination of the
tissues of 25 of the surviving control females and 11 of the non-surviving
control females for a total of 36 animals. All of the slides were
examined for each animal and the results were compared to the microscopic
reports provided by Searle Laboratories. The inconsistencies (findings
that differed from those reported by Searle) are listed below:
(64)
In most cases the inconsistencies represent findings that were not
diagnosed or reported by Searle. Copies of Searle's microscopic pathology
reports for each of the animals listed below are attached as exhibit #60.
Female Rat No. F13CF (Path. No. 95617)
Small Intestine - Diverticulum with mucosal necrosis and
cellular inflammatory infiltrate.
Female Rat No. F15CF (Path No. 95618)
Pancreas - Focal hyperplasia
Female Rat No. F16CF (Path No. 95619)
Heart - Focal Fibrosis.
Kidney - Mild chronic nephritis.
Female Rat No. H10CF (Path 95624)
Ovary - Neoplasm - probably granulosa cell tumor.
Female Rat No. H19CF (Path. No. 95626)
Kidney - Focal calcification.
Ovary - Neoplasm - probably granulosa cell tumor.
Female Rat No. H30CF (Path. No. 95628)
Kidney - Focal calcification.
Female Rat - No. K25CF (Path No. 95630)
Kidney - Focal calcification.
Female Rat No. K29CF (Path No. 95631)
Heart - Focal fibrosis
Kidney - Focal calcification
Female Rat No. M4CF (Path No. 95632)
Liver - Focal hyperplasia
Female Rat No. M10CF (Path No. 95634)
Kidney - Focal calcification.
Pituitary - Adenoma
Ovary - Fibrosis and Pigmentation.
(65)
Female Rat No. M15CF (Path No. 95635)
Pituitary - Adenoma.
Ovary - Cyst.
Female Rat No. B30CF (Path No. 95801)
Kidney - Focal calcification.
Female Rat D29CF (Path No. 95803)
Urinary Bladder (1) Chronic diffuse inflammation.
(2) Diffuse mild hyperplasia.
The second phase of the review consisted of the microscopic examination of
all tissues from the high dose females - a total of 36 animals. The
inconsistencies are listed below:
Female Rat No. B14HF (Path. No. 95657)
Eye was reported as not examined but eye was present and
normal.
Female Rat No. F25HF (Path. No. 95823)
Urinary Bladder - Mild diffuse hyperplasia.
Female Rat No. H7HF (Path No. 95623)
Ovary - Neoplasm - probably granulosa cell tumor.
Female Rat No. H9HF (Path No. 95665)
Heart - Focal fibrosis.
Urinary Bladder - Mild focal hyperplasia.
Female Rat No. H15HF (Path No. 95665)
Lymph Node - The diagnosis of lymphoma, benign, was present on
the Searle microscopic report. According to Dr. Frith, lymphoma
is generally not considered to be benign and he would diagnose
lympphosarcoma.
Female Rat NO. H18HF (Path No. 95667)
Pituitary - Adenoma.
Brain - Mild bilateral hydrocephalus.
Female Rat No. K18HF (Path No. 95824)
Pituitary - Adenoma
Female Rat No. K24HF (Path. No. 95671)
Mass noted grossly - nothing consistent with mass reported
microscopically.
(66)
Female Rat No. - M2HF (Path. No. 95672)
Uterus - Chronic mild endometritis.
Female Rat No. M30HF (Path. No. 95343)
Kidney - Focal calcification.
Uterus - Chronic mild endometritis.
Female Rat No. M30HF (Path. No. 95675)
Pancreas - Focal hyperplasia.
The third phase of this review consisted of microscopic verification of
all masses reported grossly at necropsy from all female animals not
examined in phases 1 and 2 and included a total of 73 animals. The
inconsistencies are listed below:
Female Rat No. D10Lf (Path No. 92521)
Subcutaneous mass was diagnosed as an angiofibroma on Searle
report. The lesion is more consistent with an angiosarcoma.
Female Rat No. K9MF (Path. No. 95707)
Uterus - Polyp.
Female Rat No. M1LF (Path. No. 95844)
Tissue mass seen grossly was reported as missing and not
available for microscopic examination. The tissue was
present and was a mammary fibroadenoma.
In summary, Dr. Frith reviewed:
1) All 36 high dose females (all slides) including 3 that had
been excluded from the study due to autolysis.
2) 36 (one-half) of the control females (all slides) including
1 animal that had been excluded from the study due to auto-
lysis.
3) Remaining 73 female animals with grossly observed masses.
(sufficient slides were reviewed to substantiate the masses)
4) 5 additional animals selected by the investigators (A1HM,
A9HM, A29HM, C2CM, C24HM).
(67)
The slides reviewed in the first two categories above constituted 20% of
the total animals on the study. Dr. Frith reviewed these slides blindly
and then compared his findings with the Searle microscopic reports.
According to Dr. Frith, his findings were in agreement with those of
SEarle, for the most part. In his opinion, some of the lesions that he
reported as inconsistencies were small, and might be considered
insignificant by some pathologists. Dr. Frith did feel, however, that the
ovarian neoplasms (animals H10CF, H19CF, and H7HP, and
chronic cystitis and diffuse hyperplasia (animal D29CF) should have been reported..
Dr. Frith also considered two other discrepancies to be significant. They
were:
1) The reporting of a mass (by Searle) as missing which was
actually present (MlLF).
2) The finding of a polyp of the uterus which was not diagnosed
by Searle (K9MF)
The second of the above two discrepancies assumes even more significance
in view of the following:
The Histopathologic Summary table (table 11) in Volume I of the submission
to FDA lists the following incidence of Uterine Polyps on page 87:
Incidence of Uterine Polyps
Controls Low Medium High
1 of 69 1 of 34 4 of 34 6 of 33
(1%) (3%) (12%) (18%)
The finding of one additional uterine polyp by Dr. Frith (in animal K9MF)
increases the incidence in the mid dose to 5 of 34 (15%).
On page 82 of Volume I of the submission to FDA, is the statement: "other
sporadic findings is included endometrial hyperplasia, polyp, cyst,
congestion and squamous metaplasia." The term "sporadic findings" was
used to characterize the incidence of uterine polyps, in spite of the fact
that Searle had done a statistical analysis of these findings.
__________________________________________________________________
