Cyclic adenosine 3’,5’-monophosphate (CAMP) in pregnancy body fluids during normal and abnormal pregnancy BASIL BERND
HO
YUEN,
F.R.C.S.(C)
WITTMANN,
M.D.
KATHRYN Vunrouver,
STALEY,
B.Sc.
Columbia, Canada
British
CAMP leurls in maternal urine (MU), maternal plasma (MP), and amniotic,fiuid (AF) were measured by competitive protein-binding assay in medically uncomplicated pregnancies (Group A) and some abnormal pregnancies (Group B). In Group A, MU values increased from 3.43 + 0.3 p’Moles per gram of creatinine (mean f S.E.M.) at 12 to 29 weeks to its peak value, 5.72 ? 1.03 at 33 to 35 weeks (p < 0.025). Thereafter, th values decreased until term. In AF, CAMP levels increased from 9.4 f 1.02 at IO to I9 weeks to 32.6 ? 2.7 picomoles per milliliter between 35 and 42 u1eek.s(p < 0.005). .Vlenn +- S.E.,M. MP levels between 4 to 20 and 21 to 42 weeks showed no sign$cant df,ference (14.97 ? 0.59 and 15.89 ? 0.57 picomoles per milliliter, respectively). I~mbiliral (or-d plasma contained higher concentrations than in MP and AF. In Group B, athrrortnal ,\JU patterns were observed in hypertensive disorders, chronic renal disease, a~1 postmatuv-ity. In a single patient with hyperthyroidism, abnormal MU and AF (but rrot :ZIP) 1ez~l.s zsere obsertled. Abnormalities in MU and AF concentrations of the r~u&otide are thus not specffic for otw disease entity.
RECENT STUDIES demonstrated that the maternal urinary excretion and amniotic fluid concentrations of CAMP increase during normal pregnancy,‘, * whereas maternal and umbilical cord plasma CAMP levels have not been well documented. Abnormal amniotic fluid levels and maternal urinary excretion of the nucleotide have been reported in hypertensive disorders associated with pregnancy. 3, ’ The present study was undertaken in an attempt to confirm these findings and extend the observations to include the maternal plasma in subjects with medically uncomplicated pregnancies. Preliminary data from the umbilical cord plasma of
From thu Department University of British H o.$tpital.
three neonates are included in this report. Additionally, two patients with pre-eclampsia and a patient each with chronic renal disease, post-term pregnancy, and hyperthyroidism were studied to determine possible deviations from normal in these disorders during pregnancy.
Materials and methods Group A patients. Maternal plasma (MP), maternal urinary (MU), and amniotic fluid (AF) samples were obtained in a cross-sectional manner from medically uncomplicated pregnancies of known gestational ages. Two patients (J. G. and E. T.) were studied longitudinally; umbilical cord plasma was obtained from both infants at delivery. In a third patient (A. H.), MP, umbilical cord plasma, and AF were obtained at the time of repeat elective cesarean section for fetopelvic disproportion. Collection of samples. MP samples were obtained from an antecubital vein between 09 : 00 and 15 : 00 hours. Blood was drawn into tubes containing 0.004 to 0.006M ethylene diamine tetra-acetate (EDTA) as anticoagulant and phosphodiesterase inhibitor.5 Samples were centrifuged at room temperature within 1
of Obstetrics and Gynaecology, Columbia, Vancouver General
T-his stud? supported b a grant from The Vancouver Foundation, and in part by the UBC-VGH Technique Drz~rlopmrrzt Fund. Presented @ The nt the Thirty-first Obstetricians artd .4lberta, Canada,
Canadian Investigators in Reproduction Annual Meeting of The Society of Gynaecologists of Canada, Jasper, June 19-22, 1975.
Reprint requests: Dr. Basil Ho Yuen, Department of Obstetrics and Gynaecology, Vancouver General Hospital, Vnwouver, B. C., Canada, V5Z lM9.
597
598
Yuen,
s. hf.
Wittmann,
2‘1
and Staley
fI~tlro~hlt~rotl~ia~itlt
ARhI and ox\-twin: v;r,+al tlt.llvct-\
?Ix
(first pg.) R. 8.
26 (third peg.)
Xl. Xl.
I ‘J (first p“‘g.1
ti. L. N.
28 (third pT.cg.)
A. P.
3x (fourth PWG.)
hour of c~ollccriott. Studies in our laboraror\ :111tl b\ others” twcaled that c A51J’ x ti\it\ in rhew samples rctnained stable up to 20 hcru1-s ‘II 1‘00111 tc3tlpel‘;lntl‘~. EXghty-eight samplc~ \\‘t’r~’ ob~aitwtl f’ror~t .i(i paticttth. I‘wnt\-four hour urittc satt~plcc wrv c-ollec-WI with ;I request that rhc paticttt wstric-t htnoking and the irtcakc of c,;lffeitle-c-ontainit~~ b~vcrag~s. Forty-eight samples wet-e obtained from 19 patients. .4F samples \\crc cwllectetl bv the merhods itttlic atctl on Fig. L’. .it ttiidpregt~atic~ ( IO I() 20 \WkS). 14 .-IF s;ltllplcY \\‘cr< obtained I,1 atiitiiocetiteais priot, to inductiott ot abortion \vith h) pcrtonic- wlinr tar l)svchiatric indicx lions in tiieclicallv health\ pregnant \votnrti. In late pregnancv (3.5 to -I:! weks). 13 AF samples were obtained from tnedicall~ ttticoml~lic~~tetl pregnaucies. f’rcc ot ttletx~tiiuttt or blootl ‘fhc AF samples \j’ct‘c cotltatninatioli and NYI-c ltattdlcd ~sacrl\- like the VI’ samples. c.4MP activit\ itt thew .AI; san1ple.s. as fot lhc XIP. was stable for 3) hot1r.s at roots tCt~~~)t:ri~t~Ir~. Group B patients. I‘hew wt-r patients \vith pregnaticies of kno\vn gestational ages complicated b! Lhe medical c~cmditiona showtt in ‘I.ablc I. ‘I‘h\ roitl f‘unctiotl itt K. I>. N. \vas evaluated clitlicall\ attd biochct~~icallv b\ tlic Effecti\-c I‘hyroxitic Katie.: >IC samples from (;roup B patients and 51P ,und AF satnples frotn I*;. L.. N. [vet-p collcctctl as tlcscribcd for (~rotip .A patirttt 5. Ctiless itidicatrtl. all lxllicnts in the stud! \\.etx. fret of tiiedicatiotr except for routitw prenatal iron liw at least I’, weeks prior to coll~wioti of’ 5atiiples. :111 supernaratits (~1. plasltta. .\F. antI the filterctl
xx0
alicluots 01 211’ \\vre sroretl assayed within 1 monrh. .i\.\/U
cd
c:i.\fP.
Plastlla
at leas thatt at1d
AF
-20”
(:. ;itt(l
sat11pics
1, C’l‘(’
preparc~l for ;~a\~ after the addition of four \~olutn~s of cold rthattol ro 0.3 to 1.O ml. of. s;~ttq~l~s.~ Aftetmixing. the samples wcrc centrifugrd (approximatcl~ L’.OOO x g) at t” (1. The dried supcrnatallfs \vcw thrtt tlissolvetl in .X)0 ~1 ofassa\ buffer (-I‘ris-El) 1’4. O.O.i\l Tris with 0.004M EDTA-pH 7.5); 50 ~1 aliquots tverc assayed for cAMP. Urine samples were assayed directI\ after diluting IO0 ~1 aliquots, SO- to lOO-fold with assal buffer. .I‘hc cotnpelitivc l”‘oleirl-bindill~ assav tlcscribetl 1)~ l’cnw, Oldhatn. and \l’helan” was cmplo~ctl \\ith the c,i1LlP assa\- kit. I‘KK -1-S” (Atnershatn Searlc. Le. 5. &I.). I‘his assaL i\ highly specific. fi)r (ALIP. Sigttificant interkr~ttcc IH ‘\I‘P. cyclic guanositw tnonophosphate. 1,hosphotii~stcrasc inhibitors, bi\xlent metal ions, high salr and high buffer c~oncclt~ra~ions. as wll as Huoritle ions. ltavc bern escludctl.” .%ri;tl dilution5 of plasma and .AJ; produce inhibition c urns parallel to those obtained with CAMP standards. In rubrs contaitting cARIP free serum and AF, prepared I,\ incubatiott with beef heart phosphodiesterase S’,li’-cyclic IXcleotide,* binding of tritiated cAMP was the same as in “Sigma 1.01 Ko. l2C.K77.319. ~~H.o units of rhv l~ophiliwti enzyme were dissolved in 600 111of distilled water. ‘Three IO0 4 aliquots (total of one unit) were added to 1.5 ml. of serunl (or AF) at 4 hourly intervals. The mixture was incubated at roonl temperature with continuous stirring for 30 hours.
Volume Number
CAMP in pregnancy body fluids
125 5
Table II. Concentration (picomoles per milliliter)
of CAMP in maternal
32T
plasma
3028-
=
Weeksofgestation*
Me.an S.E.M. No *Difference between significant p > 0.1,
599
4-20
21-42
14.97 0.59 38
15.89 0.57 50
4 to 20 and
21 to 42 weeks
is not
26-
.
3p 24-22s 203‘L 18;y g 12B = IOab-
.
.
.
.
. .*~:~.‘......,, . . t .
-
.-
. ..
.‘:
-a. . *.. y: .: :. - .
*
.: *
4-
Table III. Concentration of CAMP (picomoles milliliter) in amniotic fluid
2-
per
0
I
I
I
8
4
1
16
12
Weeks
I
I
I
I
20
24
28
32
I 36
I1 40
42
of Gestation
Week.5of gestation * Fig. IO-20 Mean S.E.M. No
32.6 2.7 13
9.4 1.02 14
*Difference between significant p < 0.005.
10 to 20
1. Plasma
concentration
of CAMP
and
35
to 42
.
48
weeks
is
44 Method
of fluid
.
in the
Division
of Clinical
University
Chemistry,
.
.
I-
.
88 ,-
.
I-
. . .
,_
. . . .
Hospital.
labor
.
I-
Columbia,
0
Amniocentesis at co*rclrean section
l
Depart-
of British
collection
Amniocentesis
o During
control tubes without unlabeled CAMP. Recovery of known amounts of “cold” CAMP added to aliquots of plasma, AF, and urine further confirmed the specificity of the assay. Repeated assay of pools of plasma, AF, and urine reveal mean between assay CV of less than 12.5 per cent and mean within assay CV of less than 5 per cent. All samples were assayed in duplicate and the standard curve run in triplicate. Assay results were calculated on a programmed desk-top computer (Hewlett-Packard 9810). Data were analyzed by Student’s t test. Urinary creatinine was measured by a modified Jaffe ment of Pathology, Vancouver General
pregnancy.
50
40
reaction
in normal
35-42
.
’
. .
L-
. .
Results Group A patients. MP cA,bfP concentrations. MP data are shown in Fig. 1 and Table II. Mean 2 S.E.M. between 4 to 20 and 21 to 42 weeks were 14.97 t 0.59 and 15.89 ? 0.57 picomoles, respectively, without a significant difference (p > 0.1). UmDilid cordplasma CAMP concentrations. In subject J. C;.. neonatal
milliliter plasma,
cord
(Fig. 31
plasma
3), about
picomoles
per
labor and the highest cord
plasma
picomolrs
CAMP per
CAMP
was 61 picomoles
twofold milliliter
that
per
of maternal
obtained
during
MP level in these studies. In ET, concentration
milliliter
(Fig.
was greater 4),
several-fold
than higher
80
I a
I 12
I 16
I 20 Weeks
Fig. 2. Amniotic
fluid
I 24
I 28
I
32
I
36
I
1
40 42
of Gestation
CAMP
concentrations
in
normal
prednancy.
than levels obtained in MP during pregnancy. In AH, simultaneous MP, AF, and cord plasma obtained at the time of repeat elective cesarean section revealed levels of 17, 23, and 46 picomoles per milliliter, respectively. AF CAMP. AF data are shown in Table III and Fig. 2. Mean & S.E.M. CAMP levels increased from 9.4 + 1.02 at 10 to 20 weeks to 32.6 ? 2.7 picomoles per
600
Yuen, Wittmann, and
Cord
Plasma
Staley
Plasma
CAMP
CAMP
60.7
Cord
picomoies/mf
Plasma
CAMP
concentrations
Urinary
Urinary
CAMP
PP2
28
32 Weeks
CAMP
excretion
Excretion Postpartum
24
> 80 picomoles/ml
36
40
q
2nd Day
42
of Gestation
milliliter by 35 to 42 weeks. -1‘he difference between early and late pregnancy mean values was highh significant (p < 0.005). .MI! cz4.VIP. 51~1 data are shot\ II in -l-able II’ and Fig. 3. Mean f S.E.M. of MU expressed as micromoles per
HO
YUEN,
F.R.C.S.(C)
WITTMANN,
M.D.
KATHRYN Vunrouver,
STALEY,
B.Sc.
Columbia, Canada
British
CAMP leurls in maternal urine (MU), maternal plasma (MP), and amniotic,fiuid (AF) were measured by competitive protein-binding assay in medically uncomplicated pregnancies (Group A) and some abnormal pregnancies (Group B). In Group A, MU values increased from 3.43 + 0.3 p’Moles per gram of creatinine (mean f S.E.M.) at 12 to 29 weeks to its peak value, 5.72 ? 1.03 at 33 to 35 weeks (p < 0.025). Thereafter, th values decreased until term. In AF, CAMP levels increased from 9.4 f 1.02 at IO to I9 weeks to 32.6 ? 2.7 picomoles per milliliter between 35 and 42 u1eek.s(p < 0.005). .Vlenn +- S.E.,M. MP levels between 4 to 20 and 21 to 42 weeks showed no sign$cant df,ference (14.97 ? 0.59 and 15.89 ? 0.57 picomoles per milliliter, respectively). I~mbiliral (or-d plasma contained higher concentrations than in MP and AF. In Group B, athrrortnal ,\JU patterns were observed in hypertensive disorders, chronic renal disease, a~1 postmatuv-ity. In a single patient with hyperthyroidism, abnormal MU and AF (but rrot :ZIP) 1ez~l.s zsere obsertled. Abnormalities in MU and AF concentrations of the r~u&otide are thus not specffic for otw disease entity.
RECENT STUDIES demonstrated that the maternal urinary excretion and amniotic fluid concentrations of CAMP increase during normal pregnancy,‘, * whereas maternal and umbilical cord plasma CAMP levels have not been well documented. Abnormal amniotic fluid levels and maternal urinary excretion of the nucleotide have been reported in hypertensive disorders associated with pregnancy. 3, ’ The present study was undertaken in an attempt to confirm these findings and extend the observations to include the maternal plasma in subjects with medically uncomplicated pregnancies. Preliminary data from the umbilical cord plasma of
From thu Department University of British H o.$tpital.
three neonates are included in this report. Additionally, two patients with pre-eclampsia and a patient each with chronic renal disease, post-term pregnancy, and hyperthyroidism were studied to determine possible deviations from normal in these disorders during pregnancy.
Materials and methods Group A patients. Maternal plasma (MP), maternal urinary (MU), and amniotic fluid (AF) samples were obtained in a cross-sectional manner from medically uncomplicated pregnancies of known gestational ages. Two patients (J. G. and E. T.) were studied longitudinally; umbilical cord plasma was obtained from both infants at delivery. In a third patient (A. H.), MP, umbilical cord plasma, and AF were obtained at the time of repeat elective cesarean section for fetopelvic disproportion. Collection of samples. MP samples were obtained from an antecubital vein between 09 : 00 and 15 : 00 hours. Blood was drawn into tubes containing 0.004 to 0.006M ethylene diamine tetra-acetate (EDTA) as anticoagulant and phosphodiesterase inhibitor.5 Samples were centrifuged at room temperature within 1
of Obstetrics and Gynaecology, Columbia, Vancouver General
T-his stud? supported b a grant from The Vancouver Foundation, and in part by the UBC-VGH Technique Drz~rlopmrrzt Fund. Presented @ The nt the Thirty-first Obstetricians artd .4lberta, Canada,
Canadian Investigators in Reproduction Annual Meeting of The Society of Gynaecologists of Canada, Jasper, June 19-22, 1975.
Reprint requests: Dr. Basil Ho Yuen, Department of Obstetrics and Gynaecology, Vancouver General Hospital, Vnwouver, B. C., Canada, V5Z lM9.
597
598
Yuen,
s. hf.
Wittmann,
2‘1
and Staley
fI~tlro~hlt~rotl~ia~itlt
ARhI and ox\-twin: v;r,+al tlt.llvct-\
?Ix
(first pg.) R. 8.
26 (third peg.)
Xl. Xl.
I ‘J (first p“‘g.1
ti. L. N.
28 (third pT.cg.)
A. P.
3x (fourth PWG.)
hour of c~ollccriott. Studies in our laboraror\ :111tl b\ others” twcaled that c A51J’ x ti\it\ in rhew samples rctnained stable up to 20 hcru1-s ‘II 1‘00111 tc3tlpel‘;lntl‘~. EXghty-eight samplc~ \\‘t’r~’ ob~aitwtl f’ror~t .i(i paticttth. I‘wnt\-four hour urittc satt~plcc wrv c-ollec-WI with ;I request that rhc paticttt wstric-t htnoking and the irtcakc of c,;lffeitle-c-ontainit~~ b~vcrag~s. Forty-eight samples wet-e obtained from 19 patients. .4F samples \\crc cwllectetl bv the merhods itttlic atctl on Fig. L’. .it ttiidpregt~atic~ ( IO I() 20 \WkS). 14 .-IF s;ltllplcY \\‘cr< obtained I,1 atiitiiocetiteais priot, to inductiott ot abortion \vith h) pcrtonic- wlinr tar l)svchiatric indicx lions in tiieclicallv health\ pregnant \votnrti. In late pregnancv (3.5 to -I:! weks). 13 AF samples were obtained from tnedicall~ ttticoml~lic~~tetl pregnaucies. f’rcc ot ttletx~tiiuttt or blootl ‘fhc AF samples \j’ct‘c cotltatninatioli and NYI-c ltattdlcd ~sacrl\- like the VI’ samples. c.4MP activit\ itt thew .AI; san1ple.s. as fot lhc XIP. was stable for 3) hot1r.s at roots tCt~~~)t:ri~t~Ir~. Group B patients. I‘hew wt-r patients \vith pregnaticies of kno\vn gestational ages complicated b! Lhe medical c~cmditiona showtt in ‘I.ablc I. ‘I‘h\ roitl f‘unctiotl itt K. I>. N. \vas evaluated clitlicall\ attd biochct~~icallv b\ tlic Effecti\-c I‘hyroxitic Katie.: >IC samples from (;roup B patients and 51P ,und AF satnples frotn I*;. L.. N. [vet-p collcctctl as tlcscribcd for (~rotip .A patirttt 5. Ctiless itidicatrtl. all lxllicnts in the stud! \\.etx. fret of tiiedicatiotr except for routitw prenatal iron liw at least I’, weeks prior to coll~wioti of’ 5atiiples. :111 supernaratits (~1. plasltta. .\F. antI the filterctl
xx0
alicluots 01 211’ \\vre sroretl assayed within 1 monrh. .i\.\/U
cd
c:i.\fP.
Plastlla
at leas thatt at1d
AF
-20”
(:. ;itt(l
sat11pics
1, C’l‘(’
preparc~l for ;~a\~ after the addition of four \~olutn~s of cold rthattol ro 0.3 to 1.O ml. of. s;~ttq~l~s.~ Aftetmixing. the samples wcrc centrifugrd (approximatcl~ L’.OOO x g) at t” (1. The dried supcrnatallfs \vcw thrtt tlissolvetl in .X)0 ~1 ofassa\ buffer (-I‘ris-El) 1’4. O.O.i\l Tris with 0.004M EDTA-pH 7.5); 50 ~1 aliquots tverc assayed for cAMP. Urine samples were assayed directI\ after diluting IO0 ~1 aliquots, SO- to lOO-fold with assal buffer. .I‘hc cotnpelitivc l”‘oleirl-bindill~ assav tlcscribetl 1)~ l’cnw, Oldhatn. and \l’helan” was cmplo~ctl \\ith the c,i1LlP assa\- kit. I‘KK -1-S” (Atnershatn Searlc. Le. 5. &I.). I‘his assaL i\ highly specific. fi)r (ALIP. Sigttificant interkr~ttcc IH ‘\I‘P. cyclic guanositw tnonophosphate. 1,hosphotii~stcrasc inhibitors, bi\xlent metal ions, high salr and high buffer c~oncclt~ra~ions. as wll as Huoritle ions. ltavc bern escludctl.” .%ri;tl dilution5 of plasma and .AJ; produce inhibition c urns parallel to those obtained with CAMP standards. In rubrs contaitting cARIP free serum and AF, prepared I,\ incubatiott with beef heart phosphodiesterase S’,li’-cyclic IXcleotide,* binding of tritiated cAMP was the same as in “Sigma 1.01 Ko. l2C.K77.319. ~~H.o units of rhv l~ophiliwti enzyme were dissolved in 600 111of distilled water. ‘Three IO0 4 aliquots (total of one unit) were added to 1.5 ml. of serunl (or AF) at 4 hourly intervals. The mixture was incubated at roonl temperature with continuous stirring for 30 hours.
Volume Number
CAMP in pregnancy body fluids
125 5
Table II. Concentration (picomoles per milliliter)
of CAMP in maternal
32T
plasma
3028-
=
Weeksofgestation*
Me.an S.E.M. No *Difference between significant p > 0.1,
599
4-20
21-42
14.97 0.59 38
15.89 0.57 50
4 to 20 and
21 to 42 weeks
is not
26-
.
3p 24-22s 203‘L 18;y g 12B = IOab-
.
.
.
.
. .*~:~.‘......,, . . t .
-
.-
. ..
.‘:
-a. . *.. y: .: :. - .
*
.: *
4-
Table III. Concentration of CAMP (picomoles milliliter) in amniotic fluid
2-
per
0
I
I
I
8
4
1
16
12
Weeks
I
I
I
I
20
24
28
32
I 36
I1 40
42
of Gestation
Week.5of gestation * Fig. IO-20 Mean S.E.M. No
32.6 2.7 13
9.4 1.02 14
*Difference between significant p < 0.005.
10 to 20
1. Plasma
concentration
of CAMP
and
35
to 42
.
48
weeks
is
44 Method
of fluid
.
in the
Division
of Clinical
University
Chemistry,
.
.
I-
.
88 ,-
.
I-
. . .
,_
. . . .
Hospital.
labor
.
I-
Columbia,
0
Amniocentesis at co*rclrean section
l
Depart-
of British
collection
Amniocentesis
o During
control tubes without unlabeled CAMP. Recovery of known amounts of “cold” CAMP added to aliquots of plasma, AF, and urine further confirmed the specificity of the assay. Repeated assay of pools of plasma, AF, and urine reveal mean between assay CV of less than 12.5 per cent and mean within assay CV of less than 5 per cent. All samples were assayed in duplicate and the standard curve run in triplicate. Assay results were calculated on a programmed desk-top computer (Hewlett-Packard 9810). Data were analyzed by Student’s t test. Urinary creatinine was measured by a modified Jaffe ment of Pathology, Vancouver General
pregnancy.
50
40
reaction
in normal
35-42
.
’
. .
L-
. .
Results Group A patients. MP cA,bfP concentrations. MP data are shown in Fig. 1 and Table II. Mean 2 S.E.M. between 4 to 20 and 21 to 42 weeks were 14.97 t 0.59 and 15.89 ? 0.57 picomoles, respectively, without a significant difference (p > 0.1). UmDilid cordplasma CAMP concentrations. In subject J. C;.. neonatal
milliliter plasma,
cord
(Fig. 31
plasma
3), about
picomoles
per
labor and the highest cord
plasma
picomolrs
CAMP per
CAMP
was 61 picomoles
twofold milliliter
that
per
of maternal
obtained
during
MP level in these studies. In ET, concentration
milliliter
(Fig.
was greater 4),
several-fold
than higher
80
I a
I 12
I 16
I 20 Weeks
Fig. 2. Amniotic
fluid
I 24
I 28
I
32
I
36
I
1
40 42
of Gestation
CAMP
concentrations
in
normal
prednancy.
than levels obtained in MP during pregnancy. In AH, simultaneous MP, AF, and cord plasma obtained at the time of repeat elective cesarean section revealed levels of 17, 23, and 46 picomoles per milliliter, respectively. AF CAMP. AF data are shown in Table III and Fig. 2. Mean & S.E.M. CAMP levels increased from 9.4 + 1.02 at 10 to 20 weeks to 32.6 ? 2.7 picomoles per
600
Yuen, Wittmann, and
Cord
Plasma
Staley
Plasma
CAMP
CAMP
60.7
Cord
picomoies/mf
Plasma
CAMP
concentrations
Urinary
Urinary
CAMP
PP2
28
32 Weeks
CAMP
excretion
Excretion Postpartum
24
> 80 picomoles/ml
36
40
q
2nd Day
42
of Gestation
milliliter by 35 to 42 weeks. -1‘he difference between early and late pregnancy mean values was highh significant (p < 0.005). .MI! cz4.VIP. 51~1 data are shot\ II in -l-able II’ and Fig. 3. Mean f S.E.M. of MU expressed as micromoles per
