Changes in plasma histaminase activity during normal early human pregnancy and pregnancy disorders

Changes in plasma histaminase activity during normal early human pregnancy and pregnancy disorders

OBSTETRICS Changes in plasma histaminase activity during normal early human pregnancy and pregnancy disorders MICHAEL A. BEAVEN, PH.D. JOHN R MARSHA...

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Changes in plasma histaminase activity during normal early human pregnancy and pregnancy disorders MICHAEL A. BEAVEN, PH.D. JOHN R



A sensitit•e isotopic assay was ernplo_ved to follow the changes in plasma histaminase activitv during pregnancy in patients who become pregnant following spontaneous and induced ovulation. Values in nonpregnant women ranged from 0.2 to 2.7 pmoles of histamine deaminated per miliiiiter of plasma per hour. In pregnant women, plasma histaminase activit_v began to increase 9 to 28 days following the presumed da_v of ovulation and then rose exponentially ·with a doubling time of 4 to 5 da)'S. Peak ·values, I .500 fmwles per milliliter per hour, were rearhed during the third trimester of prrgnancy. The data suggested that the increase in histaminase activity actually started at about the same time in all patients, although the early rise in enz_vme activity was not apparent in women who had high histaminase activity before pregnancy. Plasma histaminase did not rise in patients with spontaneous abortion, blighted ovum with hydatidiform degnzeration, or choriocarcinoma. The high levels of chorionic gonadotropin and low l(zir/s of histarninase activity :St't'n in patirnf.._\ 1.1Jith trophoblastic neoplasia suggest that these tests may be of value in the diagnosis of trophoblastic neoplasia early in !Jre!TI/allrY. - ..:') -_,


of the enzyme are found in this tissue. 3 - 5 Much lower enzyme activity is found in the fetal placenta or fetal circulation. 3 • 4 Histaminase or diamine oxidase, as it is more generally referred to, oxidatively deaminates histamine and a variety of diamines, such as putrescine. 6 It has been assayed by measurement of the deamination of histamine (histaminase activity),* by biologic 2 or colorimetric 7 procedures, and by the deamination of 14 C-putrescine (diamine oxidase activity) according to procedures developed by Okuyama and Kobayashi.R Both assays give comparable values and appear to be

early pregnancy, plasma levels of the enzyme, histaminase (diamine: 0 2 oxidoreductase [deaminating] EC: increase markedly. 2 The levels reach a maximum by the seventh month of pregnancy and then remain at high levels until delivery.2 The source of the enzyn1e is thought to be the decidual portion of the placenta since very high levels DURING

From the Pu/monar)' Branch, National Heart and Lung Institute, and the Reproductive Research Branch, National Institute of Child Health and Human Drvelopmenl, National Institutes of Health.

. 5, 1974. Receivr'dfur jJubliration . "'Jovembcr Accepted Del!'mber .J, 197.J.

*In this paper the term, histaminase activity. is used to refer to enzyme activity measured with histamine as substrate and diamine oxidase activity with putrescine was used as substrate.

Reprint requests: Dr. 1'"1ichael A. Beaven, Pulmonary Branch, National Heart and Lung Institute, Bethesda, Maryland 20014.



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tion in these patient~. During ihe cuurse of these studies, disorders became apparent in several patients and the effect of these disorders on the rise in enzvmc activity was itwestigated.

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DAYS Fig. 1. Plasma histaminase activity in a subject during normal. spontaneously ovulatory cycle (upper curve) and in a second subject during an HMG/HCG-induced ovulatory cvcle (lower curve).

equally sensitive. 9 ~A.. rnicromodification of the Okuyama and Kobayashi assay~ has been used exten4 sivPiv - - / hv -; Sonthrf'n -- ·--- ;mrl -· his ;~rcnci;Jtes ---- -------- in this ------ countn· ,' . ~









and by workers in Sweden, 9 · 17 · 18 to monitor the rise in enzyme activity during pregnancy and to detect disorders related to pregnancy. such as impending abortion, 13 • 16 retention of placental fragments. and trophoblastic tumorsY· 18 • 1B The precise time at which enzyme activity starts to rise in pregnancy has not been established with certainty. Ahlmark 2 detected a rise in histaminase activity 7 weeks after the last menstrual period in all subjects and 5 weeks in one subject. Southren and co-workers 10 observed high diamine oxidase activity in a patient ti weeks after her last menstrual period. In both studies, the rise in enzyme activity was exponential and rapid. This study utilizes a sensitive tritium-release assay oJ hista1ninase assay \vhich vvas developed In thi~ laboratory 20 to follow the rise in plasma enzyme activit) in early pregnancy in patients 'vho became pregnant after induction of ovulation. This made it possible to determine the precise dates of ovulation and concep10 16 -

were nonpregnant normal \·olunteers. 19 were normal \'p]unteers \vho \Vere pregnant, six were anovulatory, infertile women who received sequential administration of human menopausal gonadotropins [HMG] and human chorionic gonadotropins [HCG] to induce ovulation. and a seventh patient received clomiphene citrate. 21 • 22 Other subjects included a patient with gestational trophoblastic neoplasia and three were men with choriocarcinoma of the testis. The cycle in one normal volunteer (Fig. 1) was presumed to be ovulatory on the basis of its length, 2H days. and the presence of premenstrual molimina. In subjects rccei\·ing HMG/HCG the cycle was presumed to be ondaton on the basis of length. BBT changes. premenstrual molimina. and plasma progesterone levels of greater than 5 ng. per milliliter during the luteal phase. Pregnancy outcome in the pregnant subjects was as foliows: i H normal volunteers carried to term. one normal volunteer aborted during the first trimester. the six patients who conceived following HMG/HCG carried to term. the patient who received clomiphene citrate aborted at 3S davs gestational age. and the patient with gestational trophoblastic neoplasia was aborted in her tv;cnt\·-sccond week b\· saline ;unniotic in fusion. 'I'irnc of O\'tdation was arbitrari)y set at the fifteenth clav following the first clay of the preceding menses in the normal \olunteers and at the day following HCC administration in the subjects receiving HMG/HCG. ln the patient rccei\ing clomiphene citrate ovulation \\·as set at the ci
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Fig. 2. Plasma histaminase activity at various times before pregnancy (open circles) and during pregnancy (closed circles) in 25 women. In six of these women, ovulation was induced by administration of HMG and HCG. Other women were normal volunteers. Day 0 indicates day of HCG ad1ninistration or the fifteenth day after the iast menstrual period.

high enzyme aclivil y. The values of histaminase activity were calculated as pmoles of histamine deaminated per milliliter.

Results Plasma histaminase activity during a normal, spontaneously ovulatory cycle and an HMG/HCG-induced ovulatory cycle are shown in Fig. l. Although there was dav-to-dav variation in histaminase activitv. values remained within a narrow range for each individual. In both subjects. luteal-phase values appear lower than follicular-phase values. Fig. 2 indicates histaminase activity in the six women who conceived on HMG/HCG therapy as well as activity in normal pregnant patients as determined in random samples obtained at various times before and during pregnancy. Values before pregnancy ranged from 0.2 to 4.1, with a mean value of 1.3 ± 0. 7 (one standard deviation) pmoles per milliliter per hour and compared with values of 1.6 ± 0.9 in nonpregnant /



"'.ron-.en (n = 22). Soon after conception, histan-.inase activity began to rise steeply and ultimately reached levels of about 50 pmoles per milliliter per hour by day 60 and 500 pmoles per milliliter per hour by day 120. Serial determination of histaminase activity in five of the six women who conceived with HMG/HCG therapy are shown in Fig. 3. Prior to pregnancy. values for each patient appeared to vary within a narrow range for the individual patients. Between-patient variability was greater than within-patient variability. Soon after conception there was a steep increase in plasma histaminase activity which appeared to be exponential, with a doubling time of 4 to 5 days. The rate of rise was similar for all patients. The point at which the rise was first apparent, however, was dependent on nonpregnant levels. Patients with low nonpregnant levels showed a rise beginning 8 to 19 days following presumed ovulation whereas, in patients with higher nonpregnant values, this rise did not occur until as late as day 28 (Fig. 3).


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0·1'--_-:s'-::o--l.---4~o:--~-_~2o-:::-"---:o:--..._-::2'=o---'-4.:-:.o:--'---:-so:::--' DAYS Fig. 3. Plasma histaminase activity in five patients before and during early pregnancy. Ovulation was induced bv sequential administration of HMG and HCG. Day 0 indicates day of HCG administration. Ovulation is assumed to have occurred on dav !.

Plasma histaminase activity Lliled to rise abme tht' normal range in a patient who conceived following induction of ovulation by administration of clomiphene but then spontaneously aborted a blighted ovum with hydatidif(>rm degent>ration (Fig. 4, A) and in a normal volunteer who conceived but then spontaneously aborted 33 days after the last menstrual period (Fig. 4, B). Likewise, plasma histaminase activitY remained less than 1.5 prnoles per milliliter per hour in a patiem with gestational trophoblastic neoplasia. Three males with testicular choriocarcinoma also had normal values (Table I).

Fig. 4 . .-I, Plasma histaminase activity during the development of a hvatidit()rlll mole. The patient received clomiphene citrate to induce ovulation. She was diagnosed as pregnant from the rise in plasma HCG and an increase in basal temperature. On day :Ei (day 0 indicates day of drug administration) a hvatidiform mole wa' expelled. B. Activity in a normal volunteer who became pregnant and aborted :~;1 days after the presumed day of ovulation (da: 0).

Table I. Plasma histaminase arti\ity rhoriocarci nt Jill a

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~In pmoles of histamine deaminated per milliliter of plasma per hour. t Dav befi>re abort.ion. n)av of abortion (saline abortion). ~Twenty-five days after abortion. "Choriocarcinoma of the lcstis. Values arc of pla-;ma samples taken "" different days over a 4 wed.; period.


The availability of patients whose ovulation has been induced with HMG and HCG has made it possible to determine the exact time at which histaminase activity begins to rise during early pregnancy. The present studies show that this increase begins between 9 and 2R days following ovulation leading to pregnancy. The early rise appears to he masked in individuals who have high histaminase activity bef(n-e pregnancy. Thus. in

actuality. ir probably begins between cla\S 9 and 16. Implantation is thought to occur on days 7 to 8. According to Hertig and Rock 23 the developing lacunae receive maternal blood on about day II. It is possible that the appearance of histaminase activitY mincides with this latter event. The increases in plasma histaminase activity and HCG concentration after conception may serve as

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Changes in plasma histaminase during pregnancy

indicators of the development of the decidua and the trophoblast, respectively. as has been suggested by others. 12 · 18 · 19 The exponential rise in histaminase activity is reminiscent of a growth curve and may be a reflection of the growth of the decidual placenta. The doubling time for the increase in the histaminase act1v1ty v:as 4 to 6 days during the first 10 'Neeks of pregnancy. The rate of increase subsequently diminished at later stages of pregnancy. Similar curves were obtained by Ahlmark 2 30 years ago and more recently by Tryding and Willert. 9. These patterns of histaminase activity are similar to those seen with human chorionic gonadotropins (HCG) in pregnancies occurring following either induced21 or spontaneous ovulation. 22 HCG concentra.. tions begin to rise about day 10 and then increase exponentially, with a doubling time of 1 to 2 days; however, the rate of increase slows earlier with HCG than with histaminase activity. The sequence of events involved in the production of HCG and histaminase may be as follows: 1 to 2 days after implantation the developing trophoblast begins to produce increasing amounts of HCG which appears in plasma by day 10. Shortly thereafter the developing maternal decidua begins to synthesize increasing levels of histaminase. Depending upon the levels of plasma histaminase activity before pregnancy, an increase in plasma histaminase becomes apparent within the next 6 to 18 clays. The utility of the plasma histaminase measurement


fm the delection of disorders of placental function was suggested as early as 1944 by Ahlmark, 2 who found that in four cases of imminent abortion, uterine bleeding was attended by a fall in plasma histaminolytic (histaminase) activity. In four patients with uterine bleeding where histaminase activity remained high, the pregnancies carried to term \Vithout further complications. Similar findings have been described by other workers. Southren and associates 12 · 16 concluded that persistently low or falling histaminase activity during pregnancies are associated with a high rate of fetal wastage. _These workers suggested that the simultaneous assay of plasma diamine oxidase and plasma HGC may be useful in the diagnosis of trophoblastic tumors, and this was illustrated in one patient with choriocarcinoma where plasma diamine oxidase showed no increase. 12 Similar results were obtained by Torok and associates 19 in 22 patients with this tumor. Only in one patient was high enzyme level found. Torok and associates 19 detected only a transient increase in plasma diamine oxidase activity between weeks 13 and 16 of molar pregnancy. Before and after this period, enzyme activity remained low. The usefulness of plasma diamine oxidase determinations in following abnormalities of pregnancy was also shown in the studies of Wiiiert. 18 The lack of increase in the cases illustrated in this study also re-emphasize the possible usefulness of histaminase and HCG mesaurements in the detection of trophoblastic neoplasia verv early in pregnancy.

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5. Gunther, R. E., and Glick, B.: J. Histochem. Cytochern. 15:431, 1967. 6. Zeiier, E. A.: Fed. Proc. 24: 766, i 965. 7. Kapeller-Adler, R.: Biochem. J. 48: 99. 1957. il. Okuyama. T.. and Kobayashi, Y.: Arch. Biochem. Biophys. 95: 242, 1961. 9. Try ding, N ., and Willert, B.: Scand. J. Clin. Lab. Invest. 22: 29, 1968. 10. Southren, A. L., Kobayashi, Y., Brenner, P., and Weingold, A. B.: J. Appl. Physiol. 20: 1048, 1965. 11. Southren, A.. L., Kobayashi, Y., Jung, W., Carmody, N. C., and Weingold, A. B.: J. Clin. Endocrinol. 26: 1005, 1966. 12. Southrcn, A. L., Kobayashi, Y., Sherman, D. H .. Levine, L.. Gordon, G., and Weingold, A. B.: AM. J. OasTET. GYNECOL. 89: 199, 1964. 13. Southren. A. L., Kobayashi, Y., Weingold, A. B., and

14. 15. 16. 17. 18. 19. 20. 21. 22.


Carmody. N. C.: AM. J. OasTET. GYNECOL. 96: 502, 1966. Southren, A. L., Kobayashi, Y., Carmody, N. C., and Weingold, A. B.: AM.J. OasTET. GYNECOL. 95:615, 1966. Southren, A. L., Weingold, A. B., Kobayashi, Y., Sherman, D. H., Grimaldi, R., and Gold, E. M.: AM. J. 0BSTET. GYNECOL. 101: 899, ]968. Southren, ~A.... L., and Lee, B. 0.: Itn. J. Fertil. 16: 24. 1971. Hansson, R .. Tryding, N., and Tornquist, A.: Acta Obsiet. Gynecol. Scand. 48: 8, 1969. Willert, B.: Obstet. Gynekol. 2: 18, 1964. Torok, E. E., Brewer, J. 1., and Dolkart. R. E.: J. Clin. Endocrinol. 30: 59, 1970. Beaven, M. A., and Jacobsen, S.: .J. Pharmacol. Exp. Ther. 176: 52, 1971. Marshall, J. R., Hammond, C. B., Ross, G. T .. Jacobsen, A., Rayford, P., and Odell, W. D.: Obstet. Gynecol. 32: 760, 196il. Mishell, D. R., Jr., Thorneycroft, I. H., Nagata, Y., Nakamura, R. M.: In Rosemberg, E., Editor: Gonadotropin Therapy in Female Infertility, A.msterdam. 1975, Exerpta Medica Foundation. Hertig, A. T., and Rock, J.: AM. J. 0BSTET. GYNECOL. 47: 149. 1944.