Rare trisomy mosaicism diagnosed in amniocytes, involving an autosome other than chromosomes 13, 18, 20, and 21: karyotype/phenotype correlations.

In order to determine the significance of trisomy mosaicism of an autosome other than chromosomes 13, 18, 20, and 21, 151 such cases diagnosed prenatally through amniocentesis were reviewed. These rare trisomy mosaicism cases include 54 from 17 cytogenetic laboratories, 34 from a previous North American mosaicism survey, and 63 from published reports. All were cases of true mosaicism with information available on pregnancy outcome, and with no evidence of biased ascertainment. There were 11 cases of 46/47, +2; 2 of 46/47, +3; 2 of 46/47, +4; 5 of 46/47, +5; 3 of 46/47, +6; 8 of 46/47, +7; 14 of 46/47, +8; 25 of 46/47, +9; 2 of 46/47, +11; 23 of 46/47, +12; 5 of 46/47, +14; 11 of 46/47, +15; 21 of 46/47, +16; 7 of 46/47, +17; 1 of 46/47, +19; and 11 of 46/47, +22. As to the risk of an abnormal outcome, the data showed a very high risk (> 60 per cent) for 46/47, +2, 46/47, +16, and 46/47, +22; a high risk (40-59 per cent) for 46/47, +5, 46/47, +9, 46/47, +14, and 46/47, +15; a moderately high risk (20-39 per cent) for 46/47, +12; a moderate risk (up to 19 per cent) for 46/47, +7 and 46/47, +7 and 46/47, +8; a low risk for 46/47, +17; and an undetermined risk, due to lack of cases, for the remaining autosomal trisomy mosaics. Most cases were evaluated at birth or at termination, so subtle abnormalities may have escaped detection and developmental retardation was not evaluated at all. Comparison of the phenotypes of prenatally diagnosed abnormal cases and postnatally diagnosed cases with the same diagnosis showed considerable concordance. Since the majority of anomalies noted are prenatally detectable with ultrasound, an ultrasound examination should be performed in all prenatally diagnosed cases. In cytogenetic confirmation studies, the data showed much higher confirmation rates in cases with abnormal outcomes than in cases with normal outcomes [81 per cent vs. 55 per cent for fibroblasts (from skin, fetal tissue, and/or cord); 88 per cent vs. 46 per cent for placental cells; 22 per cent vs. 10 per cent for blood cells]. The confirmation rate reached 85 per cent when both fibroblasts and placental tissues were studied in the same case (with trisomic cells found in one or the other, or both). Therefore, one must emphasize that both fibroblasts and placental tissues should be studied. Except for 46/47, +8 and 46/47, +9, PUBS is of limited value for prenatal diagnosis of rate trisomy mosaicism. DNA studies for UPD are suggested for certain chromosomes with established imprinting effects, such as chromosomes 7, 11, 14, and 15, and perhaps for chromosomes 2 and 16, where imprinting effects are likely.

Effect of excessive GnRH-binding substance on circulating maternal hCG in human pregnancy.

Gonadotropin-releasing hormone (GnRH) can stimulate the release of placental human chorionic gonadotropin (hCG). Thus, at the onset of these studies it was the objective to define the relationship of hCG to GnRH in the maternal circulation throughout pregnancy, focusing on early pregnancy. Blood samples were collected at 8, 10, 12, 14, 16, 28 and 36 weeks of gestation during labor and the GnRH and hCG levels were determined by radioimmunoassay. Of 39 pregnancies, a GnRH-binding substance was found in the maternal circulation of three. This GnRH-binding substance resulted in erroneous GnRH levels, due to the very high non-specific binding. In the pregnant women without this GnRH-binding substance, GnRH attained highest concentrations at 12-14 weeks. The typical peak of hCG at 8-10 weeks of gestation was observed in this group, while the group of patients having the GnRH-binding substance had significantly lower hCG levels. Each of the patients with circulating GnRH-binding substance had prior pregnancy(s) and two of the three had a prior pregnancy loss. The nature of this GnRH-binding substance was investigated using gel chromatography. After incubation of [125I]GnRH with patient plasma for 3 days this substance was shown to be of high molecular weight which was ethanol precipitable. This binding substance may therefore be an antibody, since it appears to be a high molecular weight protein requiring a number of days to bind the [125I] GnRH. This GnRH-binding substance may be of physiological importance, since the circulating hCG level was significantly less in the group of patients with this substance than in those without.

Incidence and significance of chromosome mosaicism involving an autosomal structural abnormality diagnosed prenatally through amniocentesis: a collaborative study.

Among 179,663 prenatal diagnosis cases collected from ten institutions and two publications, 555 (0.3 per cent) were diagnosed as having chromosome mosaicism. Of these, 57 (10.3 per cent) were mosaic for an autosomal structural abnormality, 28 (5 per cent) for a sex chromosome structural abnormality, and 85 (15.3 per cent) were mosaic for a marker chromosome. Ninety-five cases of prenatally diagnosed mosaicism with a structural abnormality in an autosome and a normal cell line, and with a known phenotypic outcome, were collected for karyotype-phenotype correlations through our collaboration (40 cases), a prior survey (26 cases), and published reports (29 cases). They included 13 balanced reciprocal translocations, one unbalanced reciprocal translocation, four balanced Robertsonian translocations, four unbalanced Robertsonian translocations, four inversions, 17 deletions, three ring chromosomes, 19 i(20q), seven +i(12p), six other isochromosomes, and 17 partial trisomies resulting from a duplication or other rearrangement. All cases mosaic for a balanced structural rearrangement resulted in a normal phenotype. All cases of 46/46,i(20q) resulted in normal liveborns. Five of seven cases with 46/47,+i(12p) had an abnormal phenotype compatible with Killian-Pallister syndrome. The overall risk for an abnormal outcome for a mosaic case with an unbalanced structural abnormality, excluding 46/46,i(20q) and 46/47,+i(12p), is 40.4 per cent. In the same category, the study also suggested a correlation between the percentage of abnormal cells and an abnormal phenotype. For mosaicism involving a terminal deletion, the possibility of a familial fragile site should be considered.

Characterization and purification of a placental protein that inactivates GnRH, TRH and angiotensin II.

A protein that inactivates the immunoreactivity of GnRH, TRH and angiotensin II has been isolated from human term placentae. Only in the presence of DTT, a sulphydryl agent, are OXY and SRIF also inactivated by this protein. However, it is without effect on CRF, hCS, or hCG. It also inhibits the biological activity of GnRH, i.e. its ability to stimulate pituitary LH and FSH. The ability of this protein to inactivate GnRH, TRH or angiotensin II can be inhibited by various peptidase inhibitors. Thus, we have postulated that it is a chorionic peptidase, specific for these peptides, and herein called chorionic peptidase-1 (C-ase-1). Isolation of this protein, C-ase-1, has been effected using permeation, ion exchange and affinity chromatography. As estimated by SDS-PAGE and HPLC analyses, C-ase-1 has an apparent molecular weight of 58,000. It is proposed that C-ase-1 may be an important chorionic regulator of GnRH, TRH and angiotensin II levels during pregnancy.

Gestational age-related inhibition of placental hCG, alpha hCG and steroid hormone release in vitro by a GnRH antagonist.

Human placental tissues have been shown to contain gonadotrophin-releasing hormone-(GnRH)-like activity. Thus, the effect of a potent GnRH antagonist (N-Ac-Pro1,D-p-Cl-Phe2,D-Nal(2)3,6-GnRH, obtained from Syntex Laboratories) on placental hormonal release was studied. Explant cultures of placentae of 6 to 15 weeks' gestation were studied. This GnRH antagonist did not inhibit the alpha human chorionic gonadotrophin (alpha hCG), human chorionic gonadotrophin (hCG), oestrone or oestradiol release from the six- and nine-week placental cultures, but greatly suppressed the release of these hormones in the placental cultures from 13- and 15-week gestations. Synthetic GnRH partially reversed the action of this antagonist on the hormonal releases in the 15-week placental cultures. These data demonstrate a gestational age-related action of this antagonist on placental hormonal release. Thus, a role for the endogenous GnRH-like activity of the placenta in the control of placental hormonogenesis is indicated.

GnRH effects on placental hormones during gestation. III. Prostaglandin E, prostaglandin F, and 13,14-dihydro-15-keto-prostaglandin F.

We studied the release of prostaglandin E (PGE), prostaglandin F (PGF) and 13,14-dihydro-15-keto-prostaglandin F (MPF) from explants of human placentas of different gestational ages and the effect of gonadotropin-releasing hormone (GnRH) on this release. The greatest basal release of PGE, PGF and MPF was in the cultures from 9- to 13-wk placentas, with the release on the second and third days of culture increasing 4- to 10-fold from that of the first day. In cultures from 15-wk to term placentas, the initial basal release (Day 1) of these prostaglandins was only slightly higher than in cultures from 6-wk placentas. In cultures from term placentas, the later increase with extended culture was absent or very small. Addition of synthetic GnRH to the cultures from 6- to 9-wk placentas effected no significant change in release of PGE, PGF or MPF. However, GnRH added to the cultures from 13-wk placentas effected a dose-related inhibition of these prostaglandins. After 15 wk, we observed a stimulation of these prostaglandins by GnRH that was as much as 50-fold; stimulation was highly significant in the cultures from 16- and 17-wk, as well as in those from the term placentas. These data demonstrate an action of GnRH on prostaglandin release and indicate that both the basal release of PGE, PGF and MPF and the response to GnRH are related to the gestational age of the placenta.

Differential inhibition of human placental prostaglandin release in vitro by a GnRH antagonist.

Previously, we have demonstrated that the production of prostaglandins by human placental tissue varied with gestational age. In addition, we have shown that placental prostaglandin release was affected by GnRH, and that its response was also dependent on the gestational age of the placenta. Thus, we have studied the effect of a GnRH antagonist ([N-Ac-Pro1,D-p-Cl-Phe2,D-Nal(2)3,6-LHRH, Syntex Research, Palo Alto, CA) on basal prostaglandin release from placentas of 6 to 15 weeks' gestation and found that this antagonist (1 microgram/ml) effects an inhibition of the release of prostaglandin E, prostaglandin F, and 13,14-dihydro-15-keto-prostaglandin from placentas of 13 and 15 weeks of gestation. This effect was not overridden by GnRH at 10 times the antagonist concentration in the 13-week placental cultures, but was totally reversed by GnRH (10 micrograms/ml) in the 15-week placental cultures. These data demonstrate that this GnRH antagonist can affect human placental prostaglandin production at 13 to 15 weeks of gestation and indicate that endogenous placental GnRH-like activity may exert a control over placental prostaglandin release at this gestational stage.

Gonadotropin-releasing hormone effects on placental hormones during gestation: II. Progesterone, estrone, estradiol and estriol.

The release of progesterone (P), estrone (E1), estradiol (E2) and estriol (E3) from human placental tissue in vitro was found to be related to the gestational age of the placenta. The basal release of P, E1 and E2 on Day 1 of culture was highest from placentas of early gestation (9-13 wk). The release of P then declined, reaching a nadir by 15 wk, and continued at that level. The release of E1 and E2, reached a nadir at 17 weeks, and then again increased by term. In contrast, the basal release of E3 increased with increasing gestational age of the placenta. Thus, it appears that differing factors may influence placental P, E1, E2 and E3 production. In addition, the effect of synthetic gonadotropin-releasing hormone (GnRH) on these hormonal releases was studied. The stimulation of P by GnRH was greatest in placentas of 16 and 17 wk of gestation after extended culture when the basal release of P had declined. As much as a 240-fold increase was observed on the eighth day of culture. A large stimulation of P (32-fold) was also observed in the term placental cultures. A stimulation of E1 and E2 by GnRH was observed during the initial days of culture and in mid-gestational placental cultures (16-17 wk). A stimulation of E2 only was also observed at 13-15 wk and at term. A stimulation of E3 was observed in certain individual placentas. A correlation of the P and human chorionic gonadotropin (hCG) response to GnRH stimulation was noted, as well as an inverse relation of estrogens and hCG stimulation by GnRH. These data demonstrate that steroidogenic competence of the placenta differs with gestational age and that GnRH can influence steroid release. The degree and pattern of response to GnRH varied with the gestational age of the placenta and its endocrine milieu.

Gonadotropin-releasing hormone effects on placental hormones during gestation: I. Alpha-human chorionic gonadotropin, human chorionic gonadotropin and human chorionic somatomammotropin.

The release of alpha-human chorionic gonadotropin (alpha hCG), gonadotropin human chorionic gonadotropin (hCG) and human chorionic somatomammotropin (hCS) in vitro from placentas of different gestational ages was studied. In addition, the effect of gonadotropin-releasing hormone (GnRH) on these hormonal releases, as related to the gestational age of the placenta cultured and the dose of GnRH, was determined. The basal release of alpha hCG and hCG was greatest at 9-13 wk of gestation (1000-1500 ng/mg and 250-350 ng/mg, respectively). Lowest release rates were at term (28 ng/mg and 20 ng/mg, respectively). Hormonal release declined with extended culture, except from the cultures of 13- and 15-wk placentas, in which the initially high release continued throughout the 8 days of culture. The initial release of hCS was low at 6 wk, increased to maximum rates by 15 wk, and was similar to the initial rate of release at term. Gonadotropin-releasing hormone stimulated the release of alpha hCG and hCG most dramatically in cultures of 16-wk and 17-wk placentas, where as much as a 400- and 250-fold increase, respectively, on Day 6 of culture was observed (p less than 0.0001). In term placenta cultures after 6 days in vitro, a 20-fold stimulation of alpha hCG and a 10-fold increase of hCG was effected by GnRH (p less than 0.001). The largest responses of alpha hCG and hCG to GnRH were observed when estrogen levels were low. Dose-related responses were observed in some placentas, yet in some instances, maximal effects were attained with all doses utilized in these studies (0.2 to 50 micrograms/ml). These data demonstrate that human placentas of different gestational ages have varying hormonogenic capabilities in vitro. The data also establish that synthetic GnRH is capable of stimulating alpha hCG and hCG production, but the degree and pattern of response to GnRH stimulation are related to the gestational age of the placental tissue and its time in culture. The most responsive period to exogenous GnRH stimulation of alpha hCG and hCG release was on Days 5 and 6 of culture, when basal estrogen release was very low. These data support the hypothesis that hCG release might be controlled by a chorionic GnRH stimulation and suggest that local steroid levels may modulate the hCG response to GnRH stimulation.

Immunoreactive gonadotropin-releasing hormone level in maternal circulation throughout pregnancy.

Immunoreactive gonadotropin-releasing hormone was quantitated in maternal blood. Circulating levels of gonadotropin-releasing hormone were found to be significantly higher during pregnancy than in nonpregnant cycling women. The highest concentrations of gonadotropin-releasing hormone immunoreactivity were observed in the first half of pregnancy with values at 20 to 42 weeks being significantly lower. A correlation with placental human chorionic gonadotropin-releasing hormone concentrations and maternal circulating gonadotropin-releasing hormone levels was noted. Four pregnancies that resulted in premature labor and/or delivery had very low circulating maternal gonadotropin-releasing hormone concentrations, possibly reflecting placental dysfunction in these cases.

Five generations of t(4;8)(q35;q13) leading to a case of partial 8q trisomy with consideration of potential pregnancy outcomes from translocation carriers.

We describe a female infant with partial trisomy 8q who has microphthalmia, a cleft palate, micrognathia and a heart defect. Her dysmorphogenetic features closely resemble the characteristic pattern seen in the 17 cases thus far reported in the literature. Her chromosomal defect was caused by an unbalanced translocation, inherited through her father, and found to have been transmitted through at least 5 generations. Recently developed models designed to predict the most probable mode of unbalanced segregation from the meiotic quadrivalent and the likelihood that a chromosomally unbalanced fetus will survive to term are applied to this family's translocation. Also, the frequencies of potential reproductive outcomes from carriers of this translocation generated from empiric data are considered as a requisite aid to genetic counseling.

Inhibition of hCG, alpha hCG and progesterone release from human placental tissue in vitro by a GnRH antagonist.

A dramatic suppression of hCG, alpha hCG and progesterone release from midgestation, human placentas in vitro was effected when incubated with 1 microgram/ml of an antagonist to GnRH. This inhibition of hormonal release occurred rapidly and was partially restored by the addition of GnRH. Human chorionic somatomammotropin was also suppressed, but only two days following the decline of the other hormones. These data demonstrate that an antagonist to GnRH can rapidly inhibit human placental hormone release.

Prostaglandin production by human term placentas in vitro.

The release of PGs increases with increasing incubation time and the main PG produced is different in different tissues. This could implicate the presence of an inhibitory factor in vivo. PGE release is the same in patients in labor and not in labor, and this PG is mainly produced by the villi. Villi, membranes and decidua in tissue cultures from patients not in labor release more PGF than patients in labor, the difference for the decidual tissue being strikingly large. Conversion of labelled arachidonic acid by pieces of term placenta from women in labor during short-term incubations was mainly to PGE2 and its metabolites and PGD2.

Duplication (5p13 leads to pter): prenatal diagnosis and review of the literature.

We describe a patient with a reciprocal translocation t(5,13) and her three offspring. The chromosome anomaly was ascertained after the birth of her first child, who had the cri-du-chat syndrome. Amniocentesis demonstrated the presence of a dup(5p). The anomalies affecting that fetus are described and compared with the reported phenotypes of dup(5p). The extent of clinical findings in the reported cases depends on the length of the duplicated portion. The larger the duplication (p11 leads to pter), the more pronounced are the clinical signs. Physical signs are nearly absent when 5p14 leads to ter is involved. Also, translocations affecting chromosome 5 have an increased abnormal outcome when compared to D/D Robertsonian translocations. This is the first instance of antenatal diagnosis of trisomy 5p.

Studies in human fetal endocrinology: II. LH and FSH content and concentration in the pituitary.

The luteinizing hormone (LH) and follicle-stimulating hormone (FSH) content and concentration in 67 whole fetal pituitary glands have been determined by specific radioimmunoassay. A distinct pattern was observed for LH and FSH related to fetal age and sex. In the female, a mid-gestation peak of LH and FSH was found, followed by a rapid and significant decline in both gonadotropins. There was a secondary increase in the content but not the concentration from 24 weeks' gestation to term. In the male, LH and FSH content continued to increase throughout gestation. However, the rise after 20 weeks' gestation is related to the increasing pituitary weight. These data are discussed in relation to the endocrine status of the fetal gonad and hypothalamus at comparable gestational ages.

Placental luteinizing hormone-releasing factor and its synthesis.

The synthesis of a placental luteinizing hormone-releasing factor (pLRF), which is immunologically, physiochemically, and biologically indistinguishable from synthetic LRF, was demonstrated. The incorporation of 3H-labeled leucine by human placental tissue in vitro into pLRF was determined by purification on carboxymethyl-cellulose and specific immunoprecipitation of the 3H-labeled pLRF. The specific activity of the pLRF released into the medium increased 100-fold from day 1 to day 2 of culture and attained a concentration of 2.84 microcuries per microgram. These data indicate that the pLRF that was released initially was endogenous, whereas that released subsequently reflected synthesis.