ATTACHMENT 12


EXPERIMENT I ON P. 49 + 4

PURPOSE

1. The purpose of this experiment was to determine:

a. The amount of P. 49 + 4 excreted daily through the gastrointestinal 
and genite-urinary tracts of a human subject for a period of 40-60 days 
after an intravenous injection of approximately 5.0 mg. of P. 49 + 4.

b. The quantity of P. 49 present in the circulating blood four hours 
after the injection.

c. The amount of this substance which is deposited in the patella and 
the epiphysis of the tibia approximately 96 hours after the injection.

SUBJECT

1. Fifty-three year old colored male was selected who was in an 
automobile accident and sustained the following injuries:

a. Through and through laceration of the tip of the nose and the lower 
lip and a superficial laceration of the left wrist.

b. Comminuted fracture of the left femur which was in good position.

c. Comminuted fracture of the right patella which was in poor position.

d. Transverse fracture of the right radius and ulna which was in poor 
position.

PAST HISTORY

1. The patient stated that he had always been in good health except 
for a chronic urethral discharge which he has had for 10-15 years. He 
gave no history of any kidney or liver disorders.

2. The patient had always been employed as a cement mixer.

3. There was nothing of note in his family history.

PHYSICAL EXAMINATION

1. He was a well developed, well nourished colored male with the 
above evidences of trauma.

				   1

2. The other findings upon examination were:

a. He was blind in left eye, and the left cornea showed old healed 
scars and the lens was completely obliterated by cataract formation. 
The edge of the right pupil was irregular but reacted to light and
accommodation. The lens showed incipient cataract formation.

b. The patient had marked caries and pyorrhea.

c. Examination of the chest revealed no abnormal findings. His heart 
showed no enlargement to percussion. The heart sounds were of good 
quality and regular in rate and rhythm. The BP was 115/68.

3. X-ray examination of his chest was within normal limits, but 
x-ray of his knees revealed marked atrophic and hypertrophic 
arthritis with cateochondramatosis of his left knee.

4. The laboratory findings are as noted on the accompanying chart.

5. The only comments necessary are that the specific gravity of the 
urine is uniformly lower than normal. The urines show an occasional 
trace of albumin. There is a recurrent pyuria of variable degree 
which may be on the basis of chronic gonorrhea. The blood N.P.N. 
is slightly elevated, and the area nitrogen is twice the normal 
value. All of this points to somewhat diminished kidney function.

6. The patient's hemoglobin and RBC were lower than normal, and he 
showed a mild reduction of the total WBC which might have been 
produced by the sulfonamide therapy which was administered. There
was, however, no reduction in the percentage of granulocytes.

PROCEDURE

1. 100 cc of solution containing 0.3% sodium citrate and 0.5% sodium 
chloride in distilled water was prepared and autoclaved for fifteen 
minutes at 15 pounds pressure.

2. 50 microliters of P. 49 + 4 were accurately removed from an ampule 
and delivered into a glass stoppered 25 cc sterile volumetric flask.

3. 0.25 cc of the above dilution were removed with 0.5 cc tuberculin 
syringe through a 22 gauge needle.

4. The left median cubital vein of the subject was entered with a 
sterile 20 gauge needle, and 0.25 cc of P. 49 + 4 were injected. Care 
was taken to avoid leakage. The syringe was not rinsed with blood.

5. The glass ampule was sealed and returned. The solution in the 25 cc 
volumetric flash, the syringe and the two needles were returned for 
assay, and assay and determination of the actual amount of ?. 49 + 4 
injected. The amount introduced was 4.7 micrograms instead of 5.0 
micrograms.

				2

EXPERIMENT I ON P. 49 + 4

COLLECTION OF SPECIMENS

1. The urine samples were collected at irregular intervals for the 
first four days and thereafter the total 24 hour urine output was 
collected. (There is no control period because unfortunately the 
urines were pooled before and after the injection of P. 49 + 4.)

a. The urines were stored in the refrigerator in glass-top quart 
jars with rubber seals. Five cc of 40% formaldehyde was used as a 
preservative.

b. The urine was collected in new clean urinals and the urinals were 
rinsed daily with two washings of 0.1 N HCl.

2. The stool specimens were collected for each 24 hour period.

a. The stool specimens were transferred from the bed pan into 
glass-top jars by spatula, and then the bed pan was rinsed thoroughly 
three times with 200-250 cc of water and followed by a 200 cc rinse 
of .1 HHCl. These rinsings were added to the specimen jars.

b. The jars were stored in the refrigerator and enough of the 40% 
formaldehyde solution was added to the jars containing the feces to 
cover the samples.

c. A fresh bed pan was used each week.

3. A single 4 hour blood sample (10 cc) was taken. Sodium citrate 
was used as an anti-coagulant.

4. Biopsies of the patella and the epiphysis of the tibia were 
obtained for analysis when the patient was operated upon for reduction
of the fracture.

				  3