1941;1:293-297.

Cancer Res

Charles Huggins and Clarence V. Hodges

in Metastatic Carcinoma of the Prostate
Estrogen and of Androgen Injection on Serum Phosphatases
Studies on Prostatic Cancer. I. The Effect of Castration, of

Updated version

http://cancerres.aacrjournals.org/content/1/4/293.citation
Access the most recent version of this article at:







E-mail alerts
related to this article or journal. Sign up to receive free email-alerts

Subscriptions
Reprints and

. pubs@aacr.org Department at
To order reprints of this article or to subscribe to the journal, contact the AACR Publications

Permissions

. permissions@aacr.org Department at
To request permission to re-use all or part of this article, contact the AACR Publications
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
St udi e s on Pr o s t a t i c Canc e r
I. The Effect of Castration, of Estrogen and of Androgen
Injection on Serum Phosphatases in Met ast at i c
Carcinoma of the Prostate*
Charles Huggins, M.D., and Clarence V. Hodges, M.D.
(From the Department o] Surgery, the University o] Chicago, Chicago, Illinois)
(Received for publication March 22, I94I)
Car ci noma of the prost at e gl and is pecul i arl y favor-
able for endocr i ne i nvest i gat i on since f r equent serial
observations of t he act i vi t y of phosphat ases in ser um
were f ound to pr ovi de objective indices of act i vi t y of
t he neopl asm whe n the enzymes were i ncreased i n
amount above nor mal . I n t he present paper dat a are
gi ven for the val ues of serum phosphat ases i n car-
ci noma of the prost at e and in nor mal men. We shall
demonst r at e t hat the acid phosphat ase of ser um is
reduced in met ast at i c car ci noma of the prost at e by de-
creasi ng the act i vi t y of andr ogens t hr ough cast rat i on
or estrogenic i nj ect i ons and t hat this enzyme is in-
creased by i nj ect i ng androgens. We have been unabl e
to fi nd previ ous observations i ndi cat i ng any rel at i on-
shi p of hor mones to car ci noma of t he prost at e gl and.
An enzyme capable of hydrolyzing phosphoric esters was
discovered by Grosser and Husler (4) in intestinal mucosa and
kidney. Robison (I6) found that this enzyme was particularly
high in activity in growing bone and cartilage and that its ac-
tivity was greatest at pH 9 to 9.5- This "alkaline phospha-
tase" was found by Kay (9) to be increased in the serum
in certain bone diseases including metastasis of neoplasms to
bone and later work has shown that among these conditions
is carcinoma of the prostate.
Davies (3) and Bamann and Riedel (I) discovered that
there occurs in the spleen and kidney of swine and cattle, in
addition to the alkaline phosphatase, a phosphatase with an
activity maximum at pH 4.8. An enzyme believed to be identi-
cal with this "acid phosphatase" was found by Kutscher and
Wolbergs ( I I ) to be present in very large amount in the
human prostate gland. This finding of great activity of acid
phosphatase in the prostate gland was confirmed and extended
to include prostatic cancer by Gutrnan, Sproul, and Gutman (7).
The serum of certain patients with disseminated prostatic
carcinoma was found by Gutman and Gutman (6) and Bar-
ringer and Woodard (2) to exhibit increased acid phosphatase
activity. Robinson, Gutman, and Gutman (I5) summarized the
acid phosphatase activity levels of 44 patients with carcinoma of
the prostate. They concluded that a marked rise in acid phos-
phatase in serum is associated with the appearance or spread
of roentgenologically demonstrable skeletal metastases and im-
plies dissemination of the primary tumor and thus is of un-
favorable prognostic significance.
* This investigation was aided by a grant from the Com-
mittee on Research in Problems of Sex, the National Research
Council.
METHODS AND MATERIALS
The phosphat ase act i vi t y of ser um was det er mi ned
by t he met hod of Ki ng and Ar ms t r ong ( i o ) usi ng
0.005 M di sodi um monophenyl phos phat e as subst rat e.
The buffers used were 0.05 M bar bi t al - sodi um at
p H 9.3, and o. i M S6rensen' s ci t rat e-HC1 or Wal pol e' s
0.2 N sodi um acetate-acetic aci d buffers at p H 5- Al l
serums were tested i n dupl i cat e and wer e added
di rect l y to buffer-subst rat e sol ut i ons wi t hout di l ut i on;
t hey were i ncubat ed at 37.5 ~ C. for 3 ~ mi nut es. Pre-
caut i ons were observed t hat all sol ut i ons were at t hi s
t emper at ur e before t est i ng. Bl anks wer e r un by add-
i ng t he prot ei n pr eci pi t ant to t he buffer-subst rat e solu-
t i on before addi ng serum. Col or i met r i c procedures
were carri ed out wi t h t he Evel yn phot oel ect ri c color-
i met er usi ng a 6600 A filter. The results are expressed
i n Ki ng and Ar ms t r ong uni t s, a uni t bei ng defi ned as
t hat degree of phosphat ase act i vi t y whi ch at p H 9.3
( or p H 5.0, respect i vel y) and 37.5 ~ C. wi l l l i berat e
i mgm. of phenol f r om t he specified buffer-subst rat e
sol ut i on i n one-hal f hour .
Phosphat ase det er mi nat i ons at p H 5 and 9.3 were
made on t he ser um of 4 ~ nor mal men, of 2I me n wi t h
beni gn prost at i c hyper t r ophy, and of 47 me n wi t h car-
ci noma of t he prost at e. The di agnosi s of car ci noma of
t he prost at e gl and was der i ved f r om one or mor e
of t he f ol l owi ng procedures: rectal pal pat i on, cysto-
scopic exami nat i on, t r ansur et hr al resect i on wi t h mi cro-
scopic exami nat i on, or r oent genol ogi c evi dence of
skel et al metastases. Necr opsy was obt ai ned i n 2 cases.
Al l pat i ent s had x-ray st udi es of t he bony pelvis.
Ei ght pat i ent s who had car ci noma of t he prost at e
wi t h skel et al met ast ases and wi t h moder at e or gr eat
el evat i on of acid phosphat ase of ser um val ues above
20 uni t s i n Ioo cc. were selected for i nt ensi ve st udy i n
t he hospi t al . Each pat i ent also had el evat i on of al ka-
l i ne phosphat ase i n t he ser um. Bot h of these enzymes
were det er mi ned on t he ser um 3 t i mes weekl y for
ma ny weeks. Bi l at eral cast rat i on was carri ed out i n
all. Fi ve pat i ent s were i nj ect ed wi t h stilbestrol, i mgm.
293
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
294
Canc e r Re s e ar c h
daily, or estradiol benzoate, x.66 to 3.32 mgm. daily,
before castration. Thr ee patients were injected wi t h
testosterone propionate, 1 25 mgm. daily, in sesame oil.
The phosphatases of cerebrospinal fluid in 3 cases
and of spermatocele fluid in I case were det ermi ned
concurrently wi t h those of the serum on x occasion.
In this paper, units o / e n z y me re/er to IOO CC. O/ the
specified body fluid.
In 4 normal adult male dogs, serial observations of
the phosphatases of the serum were made duri ng I
week before and 4 weeks after bilateral castration.
Castration was done under ether anesthesia.
60
50
40
3C
ACID
20
I 0
, , ,
. : . . . y . ~ : .
UNITS I0 ZO
ALKALINE
30 40 50 60 70 BO
FIG. I . - - Sc a t t e r di agr am of acid and al kal i ne phosphat ase
val ues in ser um in car ci noma of t he prost at e. Or di nat es, acid
phosphat ase; abscissae, al kal i ne phosphat ase.
RESULTS
Values in carcinoma o/ the prostate and in controls.
The values wi t h standard deviation in the control
groups were: in the serum of 4 o normal men, acid
phosphatase 3.25-+-i.37 units, alkaline phosphatase
8.I5___ 1.74 units. The upper l i mi t for acid phospha-
tase is slightly hi gher than that of Gut man and Gut-
man (6) who used citrate buffer, whereas most of our
det ermi nat i ons were made wi t h acetate buffering. The
nat ure os the buffer is not the cause of the difference
obtained, however, since det ermi nat i ons made in paral-
lel usi ng citrate and acetate buffer yielded identical
results.
The values in 47 cases of carcinoma of the prostate
wi t h and wi t hout demonst rabl e metastases are shown
in Fig. i. Of these 47 patients, 23 had acid phosphatase
bel ow 4-5 units and alkaline phosphatase below i2. 5
units; of the r emai ni ng 24 patients all had elevation
1 Th e aut hor s are i ndebt ed to Dr. Er wi n Schwenk and t he
Scher i ng Cor por at i on f or gener ous donat i ons of est radi ol ben-
zoat e and t est ost erone pr opi onat e and to Dr . J. A. Morrel l of
E. R. Squi bb & Sons f or t he stilbestrol used.
of alkaline phosphatase above 12. 5 units, and 2i had
acid phosphatase values above 5 units.
Correlation between roentgenologic evidence of
skeletal metastases and serum phosphatases.--Positive
x-ray evidence of metastasis of prostatic carcinoma to
the bony pelvis was obtained in 25 cases. Both acid
and alkaline phosphatase activities of the serum were
increased above normal in 19 cases. Al kal i ne phos-
phatase alone was increased in 2 cases while acid
phosphatase was wi t hi n normal limits, and neither
enzyme was increased in 4 cases. There was no
evidence of metastasis on x-ray exami nat i on of the
pelvis of 2o patients; in i 6 men both acid and alkaline
phosphatase were normal ; in 3 men alkaline phos-
phatase was elevated but in none of this group was
the acid phosphatase increased in amount . No deduc-
tions could be made from enzyme study of the serum
wi t h respect to the osteoplastic or osteolytic nature of
the metastatic lesion.
Elyect o/castration. Aci d phosphatase.--In each of
8 patients wi t h prostatic cancer this activity rapidly
decreased fol l owi ng bilateral orchiectomy.
Following operation, in the case of P. R., there was a fall from
26 to 5 units in 7 days (Fig. 2); in the case of J. R. a
decrease from 35 to 3 units in 9 days (Fig. 3); in the case
of O. A. there was a fall from 25 to 5 units in I2 days; in
the case of C. B. a decrease from 39 to 9 units in I2 days.
Fol l owi ng castration the decrease usually persisted
in a range slightly above or below the upper limits of
the normal values; it was mai nt ai ned at this value
wi t h slight fluctuation duri ng the period of observation
whi ch in the longest case has been i8o days.
The values observed in the case of P. R. varied from 2.5
to 7.5 units during I5o days; in the case of J. R. between 3
and 6 units in xo4 days; in the case of C. B. between 7 and 9
units in I25 days; and in M. M. the values fluctuated between
9 and I5 units per ioo cc. in i8o days.
In one patient, S. R., acid and alkaline phosphatase levels of
serum were 9o and 3o units respectively i63 days after orchi-
ectomy. No other enzyme studies were made on this man, who
was the only patien t with high enzyme levels after orchiectomy.
In one patient, J. F., with benign prostatic hypertrophy who
was subjected to castration, as previously reported by Huggins
and Stevens (8), acid and alkaline serum phosphatases were
3 and 9.25 units, respectively, 2 years after orchiectomy.
In 4 dogs, the values of the serum phosphatases were
wi t hi n the range found in normal men; bilateral
orchiectomy resulted in no essential changes in their
levels.
Al kal i ne phosphat ase. NA rise was always observed
in the value of this enzyme following castration in this
series of patients. This rise was t hen succeeded by a
decrease to or towards the normal value.
I n t he case of J. R., t he val ue at the t i me of castration was 19
uni t s. Th e ma x i mu m value, 41 units, was at t ai ned 39 clays fol -
l owi ng or chi ect omy, and at lO4 days t he val ue was 2o uni t s.
I n t he case of P. R. t he precast rat i on val ue was 22 units, t he
ma x i mu m val ue f ound was 27 uni t s IO days af t er castration, and
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
Huggins et al.--Serum Phosphatase and Prostatic Cancer 295
this value decreased to 9 units at 15o days. In the case of
C. B. the precastration value was 82 units, the maxi mum rise
to 94 units was seen 12 days following orchiectomy, and the
last value obtained was 35 units 125 days following testis
removal.
Effect o/ estrogen injection.--Estrogen wa s g i v e n
d u r i n g p e r i o d s o f 8 t o 23 d a y s ; al l p a t i e n t s d e v e l o p e d
s ma l l p a l p a b l e a r e a s of s u b a r e o l a r b r e a s t t i s s ue. T h e
r e s ul t s we r e s i mi l a r t o t h o s e o b s e r v e d f o l l o wi n g cas t r a-
t i on; n a me l y , a s h a r p d e c r e a s e o f a c i d p h o s p h a t a s e
Effect o/ androgen injection.--Testosterone p r o p i o -
n a t e c a u s e d a n i n c r e a s e o f s e r u m a c i d p h o s p h a t a s e
a b o v e t h e p r e - i n j e c t i o n l evel i n t h e s e p a t i e n t s . F o l l o w-
i n g t h e c e s s a t i o n o f i n j e c t i o n s , t h e r e wa s a d e c l i n e t o
t h e p r e l i mi n a r y l evel . I n o n e p a t i e n t t hi s d e c r e a s e wa s
s u c c e e d e d b y a ma r k e d i n c r e a s e i n a c i d p h o s p h a t a s e .
T h e r e we r e n o c h a n g e s i n a l k a l i n e p h o s p h a t a s e d u r i n g
t h e p e r i o d o f i n j e c t i o n , b u t a r i s e o c c u r r e d s u b s e q u e n t
t o t r e a t me n t .
dNITS
25
,o'
s
I I i I I I I I
BILA T~'RAL ORCHIs My 2
J ' % ' ' " ' " ' N
,
I o 2 0 oAYs ~ 40 so 60 70 s o
UNITS / I ! I ' ' ' 3
$ 0
BILATERAL O RCHIECTOMY
"l~ ~ " . . . . . . . / ~176 .
p . . . . . . ..._._,..."
2o
' o F _
I i I I , I - - . I ,, I
7 i o 0AY5 2.0 3 0 4.0 5 0
5 T I L B E S T R O L - I M G . D A I L Y
I I I
4 0 5 0 D A Y S 6 0 7 0 8 0
UNI TS
6 0
SO
4 0
3 0
2 0
1O
I I I I
TESTOSTERONE P R O P I O N A T [ 2 5 M ~ DALLY ]
TOTAL 450MG.
I I I I
10 2 0 DAYS 3 0 4 0
F1o. 2. - - The effect of castration on serum phosphatases in metastatic carcinoma of the prostate in the case of P. R. Acid
phosphatase ; alkaline phosphatase - - .
Fio. 3. - - The effect of castration on serum phosphatase values in metastatic carcinoma of the prostate in the case o~
J. R. Acid phosphatase , alkaline phosphatase
Fro. 4. ---The effect of estrogen injection, I mgm. stilbestrol for 23 days, on serum phosphatases in metastatic carcinoma
of the prostate in the case of O. A. Acid phosphatase ; alkaline phosphatase . Ordinates, units per IOO cc.
of serum; abscissae, time in days.
Fro. 5. ---The effect of androgen injection, testosterone propionate, 25 mgm. daily for x8 days, on serum phosphatases
in metastatic carcinoma of the prostate in the case of O. A. Acid phosphatase ; alkaline phosphatase . . . . .
a c c o mp a n i e d b y a n i nc r e a s e of a l k a l i n e p h o s p h a t a s e i n
t h e s e r u m.
The patient P. M. was injected with 7 mgm. stilbestrol
and 46.5 mgm. estradiol benzoate in 27 days. The acid phos-
phatase of his serum decreased from 5o to 19.5 units while
the alkaline phosphatase rose from 55 to 95.5 units. J. C.
received 8 mgm. stilbestrol in 8 days. His acid phosphatase
fell from 32 to 4-5 units while the alkaline phosphatase rose
from 27.5 to 64.5 units. O. A. received 3 ~ mgm. stilbestrol in
23 days (Fig. 4) . Acid phosphatase of his serum decreased
from 48 to 4-5 units while alkaline phosphatase increased from
23 to 41.5 units.
In the case of C. B., previously castrated, injected wi t h 25
mgrn. testosterone propionate daily for 15 days, acid phospha-
tase rose from 9 to 27 units; cessation of androgen injection was
followed by a decrease to 9 units 9 days following injection.
In the case of O. A., injected with 25 mgm. testosterone pro-
pionate daily for 18 days, acid phosphatase rose from I I units
to a high value of 48.75 units reached the day fol l owi ng comple-
tion of androgen injection. This value decreased to 23.5 units
6 days fol l owi ng cessation of androgens, whereupon it rose to
46 units in the next 27 days (Fig. 5).
Distribution o[ phosphatases in body fluids in car-
cinoma o/ t he prostate.--The p h o s p h a t a s e s we r e f o u n d
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
296 Cancer Research
to be low in cerebrospinal fluid and in spermatocele
fluid al t hough the values were concurrently hi gh in
blood serum.
In the case of P. M., acid and alkaline phosphatases of the
spinal fluid were 0.75 and o units at a time when these enzymes
in the serum were 5I and 64 units respectively. Similarly in
O. A., acid and alkaline phosphatases of spinal fluid were
o and o at a time when the findings in serum were 23 and
22 units respectively. In the case of J. C., acid and alkaline
phosphatases of spinal fluid were o and 1.5 units when these
levels in serum were I9.5 and 46 units respectively. In the
case of C. R., acid and alkaline phosphatases of spermatocele
fluid were o and 1.5 when the serum levels were 49 and 62
units, respectively.
DISCUSSION
In disseminated carcinoma of the prostate the
lability of acid phosphatase in serum as influenced by
androgen supply or wi t hdrawal indicates t hat this
mal i gnancy is influenced by the activity of male hor-
mones in the host. Wi t h respect to serum enzymes,
prostatic cancer is inhibited by decreasing androgenic
hormonal function and activated by increasing the
androgens. Moreover, our data, as yet unpublished,
show t hat inhibition is not limited to the enzyme con-
tent of serum.
Cert ai n bone diseases, as shown by Kay ( 9) , are
associated wi t h increased alkaline phosphatase activity
of serum and Roberts ( i 4) discovered t hat certain liver
diseases cause a similar increase. At the present time
elevation of alkaline phosphatase of serum is regarded
as indicating only bone or hepatic disease. There is
no evidence to show t hat hepatic damage occurs wi t h
frequency in carcinoma of the prostate while extensive
bone involvement by metastasis is often observed. It
can be assumed wi t h little risk t hat elevation of alka-
line phosphatase of serum in carcinoma of the prostate
is usually due t o bone lesions, in the absence of gross
evidence of hepatopathy.
Mar ked increases in acid phosphatase in serum have
been observed only in prostatic carcinoma. Robinson,
Gut man, and Gut man ( I 5) found slight but definite
elevations of this enzyme (t he highest 8. 7 units) in
osteitis deformans and several other diseases where the
bones were involved. The dat a in the present paper
indicate t hat carcinoma of the prostate wi t h skeletal
metastasis is sometimes present wi t h serum acid phos-
phatase values wi t hi n the normal range. However,
we found t hat when there was a mar ked rise of acid
phosphatase (above io units) disseminated prostatic
carci noma was always present.
Gut man and Gut man (6) observed t hat there was
no correlation between the height of serum acid and
alkaline phosphatases in carcinoma of the prostate, and
in general our dat a agree wi t h their findings. Thi s
lack of correlation can now be explained. The ele-
vated phosphatase levels in carci noma of the prostate
are due to the action of two different types of cells,
prostatic epithelium and osteoblasts. The activity of
prostatic epithelium (as deduced from the height of
serum acid phosphatase) in disseminated cancer of
the prostate depends at least in part on the degree of
activity of androgens present in the body, whereas
elevation of alkaline phosphatase is dependent on in-
creased osteoblastic function as a result of invasion of
the bones by t umor. It is thus apparent that increased
acid and alkaline phosphatase activities in serum may
be considered separately since they indicate activity of
different types of cells. The function of those cells
contributing to the greater part of the moiety whose
maxi mum occurs at pH 5 is dependent on androgen
levels, and can be increased or decreased by adjusting
the male sex hormone level.
Havi ng established a relationship of androgenic hor-
mones to carcinoma of the prostate, the findings in
the case of O. A. (Fi g. 5) can be interpreted. In this
man the serum acid phosphatase had been greatly
increased by androgen injections and, following cessa-
tion of androgen, the enzyme decreased for i week
when it was succeeded by an abrupt spontaneous
secondary rise. The decrease following cessation of
androgen injection may be assumed to be the result
of the depression of gonadal activity caused by andro-
gen injections as indicated by Moore and Price (x~t).
The subsequent rise is explained by postulating t hat
the cancer cells had been functionally expanded by
androgen and therefore responded in an augment ed
way to hormones produced by the recovering testis.
Castration caused a marked decrease of serum acid
phosphatase but in some cases the enzyme level did
not reach normal while in others it lay in a range at
the upper limit of normal values. Much proof has been
adduced t hat androgen is produced in extragonadal
loci especially in the suprarenal gl and. The most ready
explanation for variations in serum acid phosphatase
levels following 0rchiectomy as well as the failure of
some values to reach the normal range may be postu-
lated by assumi ng varyi ng amount s or activity of
androgens produced in ext ragonadal sources in dif-
ferent individuals. It must be pointed out however
t hat uri nary assays of hormone excretion were not done
in this study. Fr om the evidence one cannot exclude
the possibility t hat loci of metastatic carcinoma as well
as the original site mi ght affect the levels of phospha-
tase and so mai nt ai n this enzyme at an abnormally
hi gh range even t hough there were no androgens
produced in extragonadal areas. Agai nst this alterna-
tive explanation is the fact t hat atrophic prostatic tissue
contains merely traces of acid phosphatase as Gut man
and Gut man (5) found in the infantile human prostate
gl and.
None of t he phosphatases has been isolated as yet
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from
Huggins et al.--Serum Phosphatase and Prostatic Cancer 297
in a pure state. The findings of none or small amount s
of these enzymes in cerebrospinal or spermatocele
fluids show t hat the enzymes are molecules which do
not cross certain membranes readily from the blood
and are not inconsistent wi t h the evidence of Kut scher
and Pany ( 12) that phosphatase is a protein.
SUMMARY AND CONCLUSIONS
~. The content in serum of phosphatase wi t h activity
maxi mum at pH 5 in normal men and in men wi t h
benign prostatic hypert rophy was 3.25 ___~.37 Ki ng and
Armst rong units in ~oo cc. Serum phosphatase wi t h
activity maxi mum at pH 9.3 in this control group was
7. 9+2. i units in IOO cc.
2. In a gr oup of 2 5 men wi t h roentgenologic evi-
dence of metastatic carcinoma to the bony pelvis, both
acid and alkaline phosphatases were increased above
normal in 19 cases, only alkaline phosphatase was in-
creased in 2 cases, and both values were wi t hi n normal
limits in 4 cases. Metastatic carcinoma of the prostate
may be present in bones when the phosphatases of
serum are normal , but when acid phosphatase is
present in activity greater t han x o units in i oo cc., dis-
seminated prostatic cancer is present.
3. In prostatic cancer wi t h mar ked elevation of acid
phosphatase, castration or injection of large amount s
of estrogen caused a sharp reduction of this enzyme
to or towards t he normal range. Al kal i ne phosphatase
values rose following castration and t hen decreased, but
more slowly t han acid phosphatase. In certain cases,
these values reached and were mai nt ai ned in a normal
range duri ng the period of observation, 18o days,
while in other patients the values were slightly above
normal.
4. In 3 patients with prostatic cancer, androgen
injection caused a sharp rise of serum acid phosphatase.
In i case following cessation of androgen there was a
decrease of t he acid phosphatase followed by a secon-
dary spontaneous rise.
5. Acid phosphatase does not readily cross mem-
branes from t he blood, since it was present only in
traces in cerebrospinal and spermatocele fluids when
concurrent serum level was high.
6. Prostatic cancer is influenced by androgenic ac-
tivity in the body. At least wi t h respect to serum
phosphatases, disseminated carcinoma of the prostate
is inhibited by el i mi nat i ng androgens, t hr ough castra-
tion or neut ral i zat i on of their activity by estrogen
injection. Cancer of the prostate is activated by
androgen injections.
REFERENCES
1. BAMANN, E., and E. RIEDEL. Ueber das Vor kommen zwei er
durch das pH Wi r kungsopt i mum unterscheidbaren Phos-
phoesterasen in tierischen Organen. Ztschr. f. physiol.
Chem., 2: 29: 125-15o. 1934.
2. BARRINGER, B. S., and H. Q. WOODARD. Prostatic Carci noma
with Extensive Intraprostatic Calcification. Tr. Am. A.
Genito-Urin. Surgeons, 3I : 363- 369. 1938.
3. DAVIES, D. R. Phosphatase Activity of Spleen Extracts. Bio-
chem. J., 28: 529- 536. 1934.
4. GROSSER, P., and J. HUSLER. Ueber das Vor kommen einer
Glycerophosphatase in tierischen Organen. Biochem.
Ztschr., 39: 1- 5. 1912.
5. GUTMAN, A. B., and E. B. GUTMAN. "Aci d" Phosphatase
and Functional Activity of the Prostate (Man) and
Preputial Glands (Rat ). Proc. Soc. Exper. Biol. &Me d. ,
39: 529- 532 9 I938.
6. GUTMAN, A. B., and E. B. GtrTMAN. An "Aci d" Phosphatase
Occurring in the Serum of Patients with Metastasizing
Carcinoma of the Prostate Gland. J. Clin. Investigation,
I 7: 473- 478- 1938.
7. GU'rMAN, E. B., E. E. SPROUL, and A. B. GUTMAN. Signifi-
cance of Increased Phosphatase Activity at the Cite of
Osteoplastic Metastases Secondary to Carcinoma of the
Prostate Gland. Am. J. Cancer, 28: 485- 495. 1936.
8. HUGGINS, C., and R. E. STEVENS. The Effect of Castration
on Benign Hyper t r ophy of the Prostate in Man. J. Urol.,
43: 7o5- 714 9 194o.
9. KAY, H. D. Plasma Phosphatase in Osteitis Deformans and
in Other Diseases of Bone. Brit. J. Exper. Path., I O:
253-256. 1929.
lO. K1NG, E. J., and A. R. ARMSTRONG. Conveni ent Method for
Det ermi ni ng Serum and Bile Phosphatase Activity. Canad.
M. A. J., 31: 376- 381. 1934. :J
11. KUTSCHER, W., and H. WOLBERGS. Prostataphosphatase.
Ztschr. f. physiol. Chem. , 236: 237- 24o. 1935-
12. KUTSCHER, W., and J. PANS'. Prostataphosphatase. Ztschr.
f. physiol. Chem. , ~3 5 : I 6 9 - I 8 9 . 1938.
13. MOORE, C. R., and D. PRICE. Some Effects of Synthetically
Prepared Male Hor mone (Androst erone) in the Rat.
Endocrinology, 2I:313-329. 1937.
14. ROBERTS, W. M. Blood Phosphatase and van den Bergh
Reaction in Differentiation of Several Types of Jaundice.
Brit. M. J., I : 734- 738. 1933.
15. ROBINSON, J. N., E. B. GVTMAN, and A. B. GUT~AN.
Clinical Significance of Increased Serum "Aci d" Phos-
phatase in Patients wi t h Bone Metastases Secondary to
Prostatic Carcinoma. J. Urol., 42: 6o2- 618. 1939.
i 6. ROBISON, R. The Possible Significance of Hexosephosphori c
Esters in Ossification. Biochem. J., 17: 286- 293. I9~3.
Research.
on July 23, 2014. 1941 American Association for Cancer cancerres.aacrjournals.org Downloaded from