scholarly journals Detection of Chromosome 21-encoded mRNA of Placental Origin in Maternal Plasma

2003 ◽  
Vol 49 (9) ◽  
pp. 1445-1449 ◽  
Author(s):  
Cees B M Oudejans ◽  
Attie T J J Go ◽  
Allerdien Visser ◽  
Monique A M Mulders ◽  
Bart A Westerman ◽  
...  
Keyword(s):  
Down Syndrome ◽  
Placental Tissue ◽  
Rna Isolation ◽  
First Trimester ◽  
Maternal Plasma ◽  
Chromosome 21 ◽  
Placental Lactogen ◽  
Plasma Samples ◽  
Human Placental Lactogen ◽  
Dnase Treatment

Abstract Background: mRNA of placental origin (i.e., human placental lactogen and β-human chorionic gonadotropin) has been demonstrated to be easily detectable in maternal plasma. We tested whether detection of chromosome 21-encoded mRNA of placental origin is possible in maternal plasma obtained during the first trimester. Methods: Plasma samples were obtained from pregnant women between weeks 9–13 of pregnancy. RNA was isolated from 800 or 1600 μL of plasma by silica-based affinity isolation and, after on-column DNase treatment, was subjected to two-step, one-tube reverse transcription-PCR with gene specific primers. Results: Three chromosome 21-encoded genes located within the Down syndrome critical region with overexpression in trisomy 21 placentas were screened for expression in early placental tissue to select their potential use for RNA based plasma screening. One of the chromosome 21-encoded genes (LOC90625) showed strong expression in first trimester placenta similar to CSH1 (human placental lactogen) and was selected for plasma analysis. The RNA isolation assay was validated with CSH1 mRNA, which could be detected in the plasma of all women tested in weeks 9–13 of pregnancy. RNA from the chromosome 21-encoded, placentally expressed gene, LOC90625, was present in maternal first-trimester plasma and could be detected in 60% of maternal plasma samples when 800 μL of plasma was used and in 100% of samples when 1600 μL of plasma was used. Conclusion: The detection of chromosome 21-encoded mRNA of placental origin in maternal plasma during the first trimester may allow development of plasma-RNA-based strategies for prenatal prediction of Down syndrome. LOC90625 is a candidate gene for this purpose.

scholarly journals Immunoreactive adrenomedullin (AM) concentration in maternal plasma during human pregnancy and AM expression in placenta

2000 ◽  
pp. 683-687 ◽  
Author(s):  
K Kobayashi ◽  
T Kubota ◽  
T Aso ◽  
Y Hirata ◽  
T Imai ◽  
...  
Keyword(s):  
Northern Blot Analysis ◽  
Plasma Concentrations ◽  
Placental Tissue ◽  
First Trimester ◽  
Maternal Plasma ◽  
Placental Lactogen ◽  
Third Trimester ◽  
Maternal Circulation ◽  
The Third ◽  
System A

Adrenomedullin (AM) is a novel vasorelaxant peptide, isolated from human pheochromocytoma. Although AM may be involved in the regulation of the cardiovascular system, a number of other mechanisms are also involved. The present study was undertaken to confirm the presence of AM in human maternal circulation and in placental function during pregnancy. Immunoreactive (ir) AM concentrations in maternal plasma were 3.4+/-0.7fmol/ml (mean+/-s.e. m.) in the first trimester, 3.3+/-1.1fmol/ml in the second trimester, 7.3+/-2.8fmol/ml in the third trimester, 4.1+/-1.9fmol/ml in early puerperium and 3.0+/-0.4fmol/ml in non-pregnant periods; the concentration in the third trimester was significantly greater than those in other periods. Plasma concentrations of estradiol (E(2)), progesterone, human placental lactogen (hPL) and human chorionic gonadotropin (hCG) were also measured, using RIA kits. Significant correlations have been demonstrated between the concentrations of irAM and those of E(2), progesterone and hPL. We therefore examined the expression of AM within the placental tissues using immunohistochemistry and northern blot analysis in order to demonstrate a correlation between the presence of AM in the placenta and maternal plasma. Using immunohistochemistry, we detected AM in the amnion at term and the expression of AM mRNA in human placental tissues using cloned human (h) AM complementary DNA as a probe. This study demonstrates the immunoreactivity of human hAM in maternal plasma during pregnancy, and suggests that hAM in maternal plasma is generated partly from placental tissue.

Human placental lactogen is a first-trimester maternal serum marker of Down syndrome

2006 ◽  
Vol 27 (1) ◽  
pp. 1-5 ◽  
Author(s):  
Michael Christiansen ◽  
Tina Lindvig Sørensen ◽  
Bent Nørgaard-Pedersen
Keyword(s):  
Down Syndrome ◽  
First Trimester ◽  
Serum Marker ◽  
Maternal Serum ◽  
Placental Lactogen ◽  
Human Placental Lactogen

scholarly journals Plasma γ-Globin Gene Expression Suggests that Fetal Hematopoietic Cells Contribute to the Pool of Circulating Cell-Free Fetal Nucleic Acids during Pregnancy

2004 ◽  
Vol 50 (4) ◽  
pp. 689-693 ◽  
Author(s):  
Tuangsit Wataganara ◽  
Erik S LeShane ◽  
Angela Y Chen ◽  
Lynn Borgatta ◽  
Inga Peter ◽  
...  
Keyword(s):  
Nucleic Acids ◽  
Pregnant Women ◽  
Globin Gene ◽  
Hematopoietic Cells ◽  
First Trimester ◽  
Maternal Plasma ◽  
Plasma Samples ◽  
Globin Mrna ◽  
Reverse Transcription Pcr ◽  
Gapdh Mrna

Abstract Background: Reports of placental mRNA sequences in the plasma of pregnant women suggest that the placenta is the predominant source of cell-free fetal nucleic acids in maternal plasma during pregnancy. We developed an assay for γ-globin mRNA concentrations to determine whether hematopoietic cells also contribute to the pool of fetal mRNA in maternal plasma. Methods: Frozen paired plasma samples obtained from 40 women before and within 20 min after elective first-trimester termination of pregnancy (TOP) were analyzed. Fresh plasma samples from eight nonpregnant individuals were included as controls. Plasma γ-globin mRNA was measured by use of real-time reverse transcription-PCR and analyzed with gestational age. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) mRNA was used to confirm the presence of cell-free RNA in each sample. Results: γ-Globin and GAPDH mRNA sequences were detected in every plasma sample. The concentrations of both messages were significantly increased in pregnancy (P <0.01). The concentrations of γ-globin mRNA were decreased in most women after TOP, but γ-globin mRNA was increased in some patients when TOP was performed later than 9 weeks of gestation. Conclusions: γ-Globin mRNA sequences can be detected and measured in fresh and frozen plasma samples. Plasma γ-globin and GAPDH mRNA concentrations are affected by pregnancy. The increased posttermination γ-globin mRNA concentrations seen in some patients suggest that the source of this message is fetal hematopoietic cells. Further study in pregnant women after 9 weeks of gestation is necessary to evaluate the potential of γ-globin mRNA as a marker for fetomaternal hemorrhage.

Circulating levels of GH-releasing hormone and GH during human pregnancy

1990 ◽  
Vol 125 (1) ◽  
pp. 161-167 ◽  
Author(s):  
M. Mazlan ◽  
C. Spence-Jones ◽  
T. Chard ◽  
J. Landon ◽  
C. McLean
Keyword(s):  
Cord Blood ◽  
Potential Role ◽  
Maternal Plasma ◽  
Placental Lactogen ◽  
Immunoradiometric Assay ◽  
Plasma Samples ◽  
Releasing Hormone ◽  
Significant Difference ◽  
Circulating Levels

ABSTRACT To study the potential role of GH-releasing hormone (GHRH) in maintaining circulating levels of GH during pregnancy, 302 maternal plasma samples were collected from non-fasted subjects at various stages of pregnancy and assayed for GHRH using a 'two-site' immunoradiometric assay. The GH and placental lactogen levels were also determined. In addition, maternal plasma samples taken during labour, amniotic fluid and cord blood were also assayed for these hormones. Maternal plasma GHRH levels were similar to non-pregnant levels throughout gestation despite fluctuations in GH values which were always higher than non-pregnant levels. There was no significant difference between GHRH levels in maternal plasma and cord blood although high GH levels were observed in the latter. These findings suggest that peripheral GHRH levels do not play an important role in maintaining circulating GH levels during pregnancy. Journal of Endocrinology (1990) 125, 161–167

EFFECT OF IONIC ENVIRONMENT ON THE RELEASE OF HUMAN PLACENTAL LACTOGEN IN VITRO

1976 ◽  
Vol 69 (3) ◽  
pp. 349-358 ◽  
Author(s):  
V. J. CHOY ◽  
W. B. WATKINS
Keyword(s):  
Placental Tissue ◽  
Placental Lactogen ◽  
Human Placental Lactogen ◽  
Short Term ◽  
Graded Response ◽  
High K ◽  
Human Placental Tissue ◽  
High Concentrations ◽  
Free Media

SUMMARY Short-term incubation of human placental tissue in Krebs–Ringer bicarbonate buffered media with various concentrations of K+ and Ca2+ showed a graded response in human placental lactogen (HPL) release at different Ca2+ concentrations, but no effect at increased K+ concentration. Media with high Ca2+ caused an inhibition of release, while Ca2+-free media caused a stimulation in HPL release. High concentrations of Mg2+ inhibited release minimally, while Ba2+ had no effect. There was no change in HPL release when Na+ concentration was increased. La3+-Locke's solution markedly inhibited release of HPL but the significance of this effect is unknown. These results suggest that Ca2+ is not required for HPL secretion from placental tissue. It seems that HPL secretion in vitro does not follow the usual pattern where a physiological stimulus or high K+ concentration causes inward movement of calcium which couples stimulation to secretion.

scholarly journals Measurement of Allelic-Expression Ratios in Trisomy 21 Placentas by Quencher Extension of Heterozygous Samples Identified by Partially Denaturing HPLC

2008 ◽  
Vol 54 (2) ◽  
pp. 437-440 ◽  
Author(s):  
Attie T J I Go ◽  
Allerdien Visser ◽  
Ofir T Betsalel ◽  
John M G van Vugt ◽  
Marinus A Blankenstein ◽  
...  
Keyword(s):  
Real Time ◽  
Trisomy 21 ◽  
First Trimester ◽  
Maternal Plasma ◽  
Rapid Identification ◽  
Chromosome 21 ◽  
Wave System ◽  
Exon 2 ◽  
Reading Frame ◽  
Time Required

Abstract Background: Measuring the allelic ratios of placental transcripts in maternal plasma permits noninvasive prenatal detection of chromosomal aneuploidy. Current methods, however, require highly specialized equipment (MALDI-TOF), limiting the widespread implementation of this powerful RNA single-nucleotide polymorphism (SNP) strategy in routine diagnostic settings. We adapted and applied the Transgenomic WAVE System and quencher extension (QEXT) for this purpose. Methods: The expressed SNP (rs2187247) in exon 2 of the placentally expressed C21orf105 gene (chromosome 21 open reading frame 105) on chromosome 21 was tested in a trisomy 21 model system in which we obtained RNA selectively released from syncytiotrophoblasts of normal and trisomy 21 placentas during first trimester. Results: In identifying heterozygous samples, we observed an exact correspondence between sequencing results and results obtained with the WAVE System. With respect to the analysis time required, the WAVE System was superior. In addition, the real-time QEXT assay (as optimized and validated with calibration standards consisting of 262-bp C21orf105 cDNA amplicons) accurately measured allele ratios after we optimized fragment purification, concentrations of input DNA and quencher label, and calculations of reporter signals. Finally, the optimized and validated QEXT assay correctly distinguished normal placentas from trisomy 21 placentas in tests of the following clinically relevant combinations: diploid homozygous (CC), diploid heterozygous (AC), triploid homozygous (AAA), and triploid heterozygous (AAC or ACC). Conclusion: The QEXT method, which is directly adaptable to current real-time PCR equipment, along with rapid identification of informative samples with the WAVE System, may facilitate routine implementation of the RNA-SNP assay for noninvasive aneuploidy diagnostics.

CONCENTRATIONS OF PLACENTAL LACTOGEN IN CHRONICALLY CATHETERIZED EWES AND FETUSES IN LATE PREGNANCY

1980 ◽  
Vol 85 (1) ◽  
pp. 27-34 ◽  
Author(s):  
M. J. TAYLOR ◽  
G. JENKIN ◽  
J. S. ROBINSON ◽  
G. D. THORBURN ◽  
H. FRIESEN ◽  
...  
Keyword(s):  
Circadian Rhythm ◽  
Late Pregnancy ◽  
Maternal Plasma ◽  
Fetal Weight ◽  
Placental Lactogen ◽  
Plasma Samples ◽  
The Mean ◽  
The Difference

SUMMARY The concentration of ovine placental lactogen (oPL) was measured by radioimmunoassay in plasma samples from chronically catheterized ewes and their fetuses from day 110 of gestation to term (about day 145). Concentrations of oPL in the plasma of the mother and fetus were raised after surgery, and remained raised for 3–5 days after the operation. Concentrations of oPL were greatest in the fetus at days 120–124 of gestation, and then declined until delivery. Mean concentrations of oPL in the fetus in late pregnancy for single, twin and triplet pregnancies were 101±6 (s.e.m.), 100±11 and 117±59 ng/ml respectively and were not significantly different. Mean concentrations of oPL in the mother in late pregnancy for single, twin and triplet pregnancies were 718±227, 1387±160 and 1510±459 ng/ml respectively; the difference between these means was significant (P <0·05). Peak concentrations were noted at days 130–139 of gestation after which concentrations fell and were significantly lower on the day of delivery (P <0·01). Concentrations of oPL in the mother showed no circadian rhythm. The mean concentrations of oPL in maternal plasma during late pregnancy was significantly correlated to the combined fetal weight at birth (r = 0·624, P <0·01).

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