Determination of fetal RhD status by maternal plasma DNA analysis.
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Base Sequence↗
Biomedical subjects
C W Redman
Publications and source records attributed to C W Redman.
Fetal heart rate patterns in term labor vary with sex, gestational age, epidural analgesia, and fetal weight.
OBJECTIVES: Our purpose was to analyze the previously unreported effect of fetal sex on the fetal heart rate in labor and to measure its magnitude in relation to the effects of other independent clinical variables. STUDY DESIGN: The last hour of the intrapartum heart rates of 1884 term singleton fetuses collected during routine clinical monitoring over 19 months in Oxford, United Kingdom, was analyzed by computerized techniques. The records were selected for completeness and continuity until within at least 30 minutes of delivery. A subset of records from earlier in labor and a separate archive of antepartum normal term records were also examined. RESULTS: Female fetuses had significantly faster heart rates than male fetuses (P <.0001). Epidural analgesia, weight percentile (adjusted for age and sex), parity, the duration of first and second stages of labor, and a fall in umbilical arterial blood pH at birth also independently modulated the fetal heart rate (all P <.0001). The effects of these independent variables on heart rate were additive, the most important being epidural analgesia as a cause of tachycardia. The effect of fetal sex was less in the first stage, 6 to 7 hours before delivery, and was not present before the onset of labor (in another 552 pregnancies at 37 to 38 weeks). CONCLUSIONS: The fetal heart rate response of female fetuses to normal labor differs from that of male fetuses. Computerized numeric analysis of intrapartum fetal heart rate patterns will need to take into account the multiple factors that influence the fetal heart rate to identify precisely which patterns predict clinical outcome.
Adult↗
Preeclampsia: an excessive maternal inflammatory response to pregnancy.
The maternal syndrome of preeclampsia has previously been ascribed to generalized maternal endothelial cell dysfunction. In this review we suggest that the endothelial dysfunction is a part of a more generalized intravascular inflammatory reaction involving intravascular leukocytes as well as the clotting and complement systems. We provide evidence from our recent work and that of others that not only supports this proposal but indicates that such an inflammatory response is already well developed in normal pregnancy and that the differences between normal pregnancy and preeclampsia are less striking than those between the normal pregnant and nonpregnant states. From this we argue that preeclampsia arises when a universal maternal intravascular inflammatory response to pregnancy decompensates in particular cases, which may occur because either the stimulus or the maternal response is too strong. We conclude that there is no specific cause for the disorder, which can be better considered as the extreme end of the range of maternal adaptation to pregnancy. We propose that poor placentation is not the cause of preeclampsia but is a powerful predisposing factor. We predict that a single preeclampsia gene will not be found, nor will either a single specific predictive test or single preventive effective measure be devised. Aspects of the hypothesis are testable, and future work should allow its confirmation or refutation.
Endothelium, Vascular↗
Expression and subcellular localization of cyclin D1 protein in epithelial ovarian tumour cells.
The expression of cyclin D1 protein in tumour sections from 81 patients with epithelial ovarian cancer was analysed using immunohistochemistry. The tumours that overexpressed cyclin D1 in more than 10% of neoplastic cells were considered positive. Thus overexpression of cyclin D1 was observed in 72/81 (89%) of the cases examined. Protein was detected in both the nucleus and the cytoplasm in 24/81 (30%) and localized exclusively in the cytoplasm in 48/81 (59%) of the tumours. Cyclin D1 was overexpressed in both borderline and invasive tumours. There was no association between protein overexpression and tumour stage and differentiation. Furthermore, no correlation between cyclin D1 expression and clinical outcome was observed. However, in tumours overexpressing cyclin D1 (n = 72), the proportion displaying exclusively cytoplasmic localization of protein was higher in those with serous compared with non-serous histology (P = 0.004, odds ratio 4.8, 95% confidence interval 1.4-19.1). Western analysis using a monoclonal antibody to cyclin D1 identified a 36 kDa protein in homogenates from seven tumours displaying cytoplasmic only and one tumour demonstrating both nuclear and cytoplasmic immunostaining. Using restriction fragment length polymorphism polymerase chain reaction and PCR-multiplex analysis, amplification of the cyclin D1 gene (CCND1 was detected in 1/29 of the tumours demonstrating overexpression of cyclin D1 protein. We conclude that deregulation of CCND1 expression leading to both cytoplasmic and nuclear protein localization is a frequent event in ovarian cancer and occurs mainly in the absence of gene amplification.
Blotting, Western↗
Purification and characterization of a complex from placental syncytiotrophoblast microvillous membranes which inhibits the proliferation of human umbilical vein endothelial cells.
The signs of pre-eclampsia are thought to arise from maternal endothelial dysfunction caused by circulating factors of placental origin. Syncytiotrophoblast microvillous membranes (STBM) cause endothelial disruption and inhibit proliferation in vitro. Significantly increased amounts of STBM can be detected in blood from pre-eclamptic women and could contribute to endothelial dysfunction in vivo. This study purified a complex from STBM which inhibits the proliferation of cultured human endothelial cells. Integral membrane proteins were solubilized with sucrose monolaurate. Anion exchange chromatography yielded two peaks of anti-proliferative activity. Only the second peak was specific to STBM and was subjected to further separation by Sephacryl S-200 gel filtration chromatography (GFC). A single peak of specific activity eluted close to the void volume, at a position unaltered by added denaturing agents, guanidium chloride or urea. On Sephacryl S-300 GFC, two peaks were obtained of 410 and 820 kDa, with similar anti-proliferative activity and protein components (by SDS-polyacrylamide gel electrophoresis). The major protein bands were as integrins alpha5 and alpha v, dipeptidyl peptidase IV, alpha-actinin, transferrin, transferrin receptor, placental alkaline phosphatase and monoamine oxidase A.
Actinin↗
Trophoblast deportation in human pregnancy--its relevance for pre-eclampsia.
The maternal syndrome of pre-eclampsia is thought to result from endothelial cell damage caused by a circulating factor derived from the placenta. This study investigates the hypothesis that trophoblast deportation may be part of the process by which this factor enters the maternal circulation. The nature and incidence of trophoblast deportation was studied in uterine vein and peripheral blood taken from normal and pre-eclamptic women at caesarean section. Trophoblasts were enriched using immunomagnetic beads to deplete leucocytes and labelled with trophoblast-specific monoclonal antibodies. Syncytiotrophoblast, cytotrophoblast, cytotrophoblast clumps and anucleate trophoblast cells were found in uterine vein blood. Cytotrophoblast cells were found to be shed less frequently than syncytiotrophoblast and the majority were probably villous in origin. Trophoblasts were found in the uterine vein blood of normal pregnant women with higher levels in pre-eclampsia. However, trophoblasts were rarely found in the peripheral circulation. There was no correlation between trophoblast numbers and either the severity of the disease, the extent of placental pathology or the inhibitory effect of uterine and peripheral vein plasma on endothelial growth in vitro. Thus, it is speculated that increased trophoblast deportation in pre-eclampsia is secondary to the structural and functional changes occurring in the placenta, rather than directly linked with the circulating endothelial cell damaging factor in pre-eclampsia.
Adult↗
A duplication of the descending colon presenting in pregnancy.
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Journal Article↗
Audit of the views of gynaecological cancer patients regarding follow-up.
The purpose of this study was to audit patients opinions on routine follow-up and discharge from the gynaecology clinic. The setting was the gynaecology clinic of a large district general hospital and the design was a cross-sectional audit analysis using a written questionnaire. The subjects were 103 women attending the gynaecological out patients for review following previous treatment for various gynaecological cancers. Ninety patients took part in the study, of these 36 (40%) wished for 'free access', 29 (32%) wished for routine hospital follow-up, and 25 (28%) wished for discharge but with follow-up by their general practitioner. There were no statistically significant differences in choices between cancer groups. The choice of 'free access'seemed to be an important choice for women. We conclude that routine hospital follow-up of women treated for gynaecological cancer is not the desire of the majority and it appears the women may have different inherent psychological characteristics.
Journal Article↗
Do women referred for colposcopy receive adequate information from the primary care team?
The objective of this study was to assess whether women had been counselled by their primary health care team and whether or not they had received any information about colposcopy. We also asked where the best information had been obtained. The setting was the colposcopy clinic of a large district general hospital, the design of which was a cross-sectional audit using a questionnaire. The subjects were 100 women attending the colposcopy clinic. The results show that 63% of women were seen by their primary health care team before the colposcopy visit and that counselling was associated with knowledge about colposcopy (P = 0.017). However, 43% of women felt that they knew nothing about colposcopy whilst 19% of women thought that the information they had obtained was not useful. We conclude that there is scope for improving the quality, timing and provision of information for women undergoing colposcopy.
Journal Article↗
Supernatants from co-cultured endothelial cells and syncytiotrophoblast microvillous membranes activate peripheral blood leukocytes in vitro.
There is evidence for both endothelial cell and peripheral blood leukocyte (PBL) activation in pre-eclampsia. Syncytiotrophoblast microvillous membranes (STBM) are shed in greater quantities from the placenta in pre-eclampsia, disrupt cultured endothelial cells in vitro and may be the immediate cause of the maternal syndrome. The aim of this study was to determine if endothelial cells co-cultured with STBM release factors that can activate PBL in vitro. Flow cytometry was used to measure changes in intracellular free ionized calcium ([Ca2+]i), pH (pHi) and reactive oxygen species (iROS) as indices of leukocyte activation. PBL from male non-pregnant donors was exposed to supernatants from human umbilical vein endothelial cells (HUVEC) cultured with STBM. The time course of changes in [Ca2+]i, pHi and iROS was determined and compared with appropriate control measurements. The test supernatants caused significant activation of granulocytes and monocytes in terms of increases in [Ca2+]i and falls in pHi and release of iROS. Lymphocytes responded only with respect to increases in iROS. The results define a possible mechanism for the activation of PBL in pre-eclampsia, as being secondary to endothelial cell activation caused by circulating STBM shed in excess amounts from the placenta.
Calcium↗
Gestational regulation of granulocyte-colony stimulating factor receptor expression in the human placenta.
A number of cytokines and their receptors are abundantly expressed at the materno-fetal interface and are thought to have a function in the regulation of placentation. Granulocyte-colony stimulating factor (G-CSF) is expressed by stromal cells in both placental tissue and maternal decidua throughout placentation. In this study, we examined the expression of placental G-CSF receptor (G-CSFR) mRNA and protein throughout gestation by ribonuclease protection assays, Western blotting, and immunohistochemistry. The major placental form of G-CSFR mRNA, corresponding to a membrane-bound form of the protein, was present in first-trimester placental tissues; levels decreased in second- and were highest in third-trimester placental tissues. Two placental G-CSFR molecules, 120 kDa and 150 kDa, were detected in first- and third-, but not second-, trimester tissues. The level of the 150-kDa G-CSFR was greater in the third- than in first-trimester samples. These differences were irrespective of whether or not the patients had received prostaglandin E1 analogues, prostaglandin E1 analogues and oxytocin, oxytocin alone, or mifepristone before labor. We demonstrated by immunohistochemistry that interstitial cytotrophoblast in first- and second-trimester decidual tissue and cytotrophoblast in term fetal membranes express G-CSFR. These data demonstrate that the expression of specific forms of placental G-CSFR is strictly cell type- and developmental stage-specific, and they suggest that G-CSFR may be important in decidual invasion of cytotrophoblast and in trophoblast function during placentation.
Abortifacient Agents, Steroidal↗
Is the provision of information leaflets before colposcopy beneficial? A prospective randomised study.
OBJECTIVE: To assess the usefulness of a leaflet distributed to women before colposcopy designed to reduce their anxiety and psychosexual morbidity by providing information. DESIGN: Prospective randomised study. SETTING: Colposcopy clinic of a large district general hospital. SAMPLE: Two hundred consecutive women undergoing colposcopy for the first time for a cervical cytological abnormality of severity no greater than moderate dyskaryosis. METHODS: Women were randomised into one of two groups (leaflet or control). Those in the leaflet group were sent an information leaflet prior to attending the clinic. In the colposcopy clinic all the women completed a State/Trait Anxiety Inventory (StAI/TrAI) and a modified psychosexual questionnaire before undergoing colposcopy. This was repeated at the six-month follow up visit. Women in the leaflet group also completed a further questionnaire on the leaflet. MAIN OUTCOME MEASURES: Differences of anxiety and psychosexual scores between leaflet and control groups. RESULTS: The leaflet was well received. There were no statistical differences in StAI and TrAI scores between the study group and the control group at either visit, although in the whole study population StAI and TrAI scores were reduced at the second visit. The leaflet group had significantly more psychosexual problems but by the second visit, the scores had improved and the two groups were similar. When the mean differences in anxiety and psychosexual scores at the initial and second visits were compared between the groups, the reduction in negative sexual feelings and deterioration of TrAI scores experienced by the leaflet group was significant. CONCLUSIONS: This study suggests that the provision of sending an information leaflet prior to colposcopy is not beneficial in isolation. Other approaches need to be considered.
Adult↗
Maternal peripheral blood leukocytes in normal and pre-eclamptic pregnancies.
OBJECTIVE: To analyse activation of maternal peripheral blood leukocytes by flow cytometric measurements of intracellular free-ionised calcium of lymphocytes, granulocytes and monocytes, separately. DESIGN: Case-control study. SETTING: High risk pregnancy service in a regional centre. MATERIAL: Samples from 10 women with pre-eclampsia, 10 appropriately-matched women with normal pregnancy, nine multigravid normal women at mid-gestation selected as being least likely to demonstrate any tendencies towards pre-eclampsia, and 11 healthy nonpregnant women of reproductive age were studied. METHODS: Using flow cytometry, intracellular free ionised calcium ([Ca2+]i) was estimated by loading the cells with Fluo-3 and measuring the changes in fluorescence intensity induced by free ionised calcium. After the basal levels were measured, the response of phagocytes to stimulation with n-formylmethionyl-leucyl-phenylalanine (fMLP) was determined. MAIN OUTCOME MEASURES: Basal [Ca2+]i of peripheral blood leukocytes. RESULTS: Median basal [Ca2+]i was significantly increased in all three subsets of leukocytes--lymphocytes, granulocytes and monocytes in pre-eclampsia--compared with the three control groups. Samples from both groups of women with normal pregnancy did not differ from those from nonpregnant women. The peak responses of monocytes to stimulation with 10 nmol fMLP were greater in samples from pre-eclamptic women, giving evidence of priming. CONCLUSIONS: Peripheral blood leukocytes are activated in pre-eclampsia in terms of basal changes in the intracellular second messenger--free ionised calcium. Peripheral blood monocytes are primed to give greater responses after stimulation with fMLP.
Adult↗
31P magnetic resonance spectroscopy of the liver in HELLP syndrome.
OBJECTIVES: Using magnetic resonance spectroscopy (MRS) to measure phosphorus-containing metabolites in the liver, this study aimed to investigate non-invasively whether or not women with haemolysis, elevated liver enzymes and low platelets (HELLP) have detectable abnormalities of hepatic energetics. SETTING: John Radcliffe Hospital, Oxford. DESIGN: Prospective study. METHODS: After giving informed consent, patients with HELLP syndrome (n = 7) and controls with severe pre-eclampsia (n = 3), were studied by 31P MRS of the liver as soon as possible after delivery (range 2-4 days) and compared with normal nonpregnant controls (n = 6). Haematological and biochemical tests were performed serially and on the day of the MRS in all pregnant patients. RESULTS: The severity of HELLP varied as follows: peak aspartate aminotransferase (range 129-2574), peak gamma glutamyl transferase (range 28-96), peak lactate dehydrogenase (range 305-2820), nadir platelets (range 25-114), peak international normalised ratio for prothrombin time (before fresh frozen plasma) (range 0.9-1.9). One pregnancy was terminated but all others resulted in live births and all mothers made uneventful, rapid recoveries. MRS-determined relative hepatic concentrations of phosphorus-containing metabolites and absolute concentrations of adenosine triphosphate did not differ significantly between groups. One patient with the most clinically severe HELLP syndrome (by laboratory criteria) exhibited magnetic resonance spectra which showed a relative increase in phosphomonoester and an absolute decrease in hepatic adenosine triphosphate (to 62% of control). CONCLUSIONS: Enthusiasm for the conservative management of HELLP syndrome that develops remote from term has been tempered by the inability to identify patients at risk for progression to hepatic necrosis. We found that most patients with HELLP syndrome had normal liver metabolism as assessed by MRS. However, clinically severe HELLP syndrome can be associated with disturbed hepatic metabolism consistent with that seen in hepatic ischaemia and/or granulocytic infiltration of the liver.
Adolescent↗
Quantitative abnormalities of fetal DNA in maternal serum in preeclampsia.
BACKGROUND: There is much recent interest in the biologic and diagnostic implication of cell-free non-host DNA in the plasma and serum of human subjects. To determine if quantitative abnormalities of circulating non-host DNA may be associated with certain pathologic processes, we used circulating fetal DNA in preeclampsia as a model system. METHODS: We studied 20 preeclamptic women and 20 control subjects of comparable gestational age (means, 32 and 33 weeks, respectively). Male fetal DNA in maternal serum was measured using real-time quantitative PCR for the SRY gene on the Y chromosome. RESULTS: The imprecision (CV) of the assay was 2.7%. The median circulating fetal DNA was increased fivefold in 20 preeclamptic women compared with 20 control pregnant women (381 vs 76 genome-equivalents/mL, P <0.001). CONCLUSIONS: These observations suggest that preeclampsia is associated with disturbances in the liberation and/or clearance mechanisms of circulating DNA. These results also raise the possibility that measurement of circulating DNA may prove useful as a marker for the diagnosis and/or monitoring of preeclampsia.
Case-Control Studies↗
Prenatal diagnosis of fetal RhD status by molecular analysis of maternal plasma.
BACKGROUND: The ability to determine fetal RhD Status noninvasively is useful in the treatment of RhD-sensitized pregnant women whose partners are heterozygous for the RhD gene. The recent demonstration of fetal DNA in maternal plasma raises the possibility that fetal RhD genotyping may be possible with the use of maternal plasma. METHODS: We studied 57 RhD-negative pregnant women and their singleton fetuses. DNA extracted from maternal plasma was analyzed for the RhD gene with a fluorescence-based polymerase-chain-reaction (PCR) test sensitive enough to detect the RhD gene in a single cell. Fetal RhD status was determined directly by serologic analysis of cord blood or PCR analysis of amniotic fluid. RESULTS: Among the 57 RhD-negative women, 12 were in their first trimester of pregnancy, 30 were in their second trimester, and 15 were in their third trimester. Thirty-nine fetuses were RhD-positive, and 18 were RhD-negative. In the samples obtained from women in their second or third trimester of pregnancy, the results of RhD PCR analysis of maternal plasma DNA were completely concordant with the results of serologic analysis. Among the maternal plasma samples collected in the first trimester, 2 contained no RhD DNA, but the fetuses were RhD-positive; the results in the other 10 samples were concordant (7 were RhD-positive, and 3 RhD-negative). CONCLUSIONS: Noninvasive fetal RhD genotyping can be performed rapidly and reliably with the use of maternal plasma beginning in the second trimester of pregnancy.
Blood Grouping and Crossmatching↗
Vascular smooth muscle oxygen consumption is reversibly stimulated by sera from women with preeclampsia.
OBJECTIVE: Preeclampsia is a complication of pregnancy that causes maternal vasoconstriction and hypertension. The disease may progress to eclampsia, which is thought to be related to cerebral vasospasm. Although there is evidence for more than one circulating factor that causes endothelial cell dysfunction in preeclampsia, little work has focused on the possibility that vascular smooth muscle function might be directly stimulated by a circulating factor. The aim of this study was to determine whether such a factor or factors could be detected by the vessels. STUDY DESIGN: Excessive vascular smooth muscle oxygen consumption was used as a screen for metabolic stimulation because pathologic arterial constriction would require oxidative metabolism to generate adenosine triphosphate. De-endothelialized porcine carotid artery (a well-validated model of human arterial contractile function) was exposed to sera from patients with preeclampsia (1:30 dilution) in a sealed chamber with an oxygen electrode, and the rate of oxygen consumption by the tissue was measured. Comparisons with the effects of sera from matched normal pregnant patients and from nonpregnant women were made. RESULTS: Exposure of vascular smooth muscle to sera from women with preeclampsia for 90 minutes resulted in greater oxygen consumption by the tissue (0.66 +/- 0.16 micromol O2 /min per gram of dry weight) than did exposure to sera of matched pregnant and nonpregnant control subjects (0.34 +/- 0.08 micromol O2 /min per gram of dry weight, P <.001, and 0.29 +/- 0.03 micromol O2 /min per gram of dry weight, P <.001, respectively). This stimulation was completely reversed by rinsing. CONCLUSIONS: There is a factor in the circulation of women with preeclampsia that has the reversible effect on vascular smooth muscle of accelerating oxygen consumption. We discuss the implications of this observation in terms of known aspects of vascular smooth muscle contractile function.
Animals↗
Serum from preeclamptic women induces vascular cell adhesion molecule-1 expression on human endothelial cells in vitro: a possible role of increased circulating levels of free fatty acids.
OBJECTIVE: The object was to determine whether serum from preeclamptic women induces expression of vascular cell adhesion molecule-1 on cultured endothelial cells. STUDY DESIGN: Endothelial cells were incubated with medium containing 20% serum (volume/volume) either from women with preeclampsia (n = 15) or from women with normal pregnancies (n = 15) matched for maternal age, gestational age, and parity. A further matched set of samples (n = 10) was exposed to endothelial cells that had previously been incubated in the presence or absence of vitamin E (40 micromol/L final concentration). Free fatty acids were determined in each sample. A mixture of free fatty acids (linoleic, oleic, and palmitic acids, 1:1:1) was added to serum from control subjects in increasing concentrations (70-280 micromol/L final concentration) to emulate preeclamptic serum and the preparation was exposed to endothelial cells. In each experiment vascular cell adhesion molecule-1 expression was determined after 16 hours of exposure by an enzyme-linked immunosorbent assay technique performed on the cell monolayer. RESULTS: Preeclamptic serum had higher levels of free fatty acids than did that of control subjects (0.71 mmol/L, 95% confidence level 0.5-0.93, vs 0.36 mmol/L, 95% confidence level 0.28-0.43). There was a statistically significant increase in vascular cell adhesion molecule-1 expression on the endothelial cells exposed to preeclamptic serum compared with those exposed to control serum (optical density 0.17 vs 0.11). Vitamin E reduced the vascular cell adhesion molecule-1 expression of endothelial cells exposed both to preeclamptic and to control serum samples in a nonspecific manner. Addition of free fatty acids to normal pregnancy serum to mimic the effect of preeclampsia resulted in increased expression of vascular cell adhesion molecule-1 on the cells. CONCLUSION: Preeclamptic serum induces vascular cell adhesion molecule-1 expression on human endothelial cells in vitro, an effect also produced by fatty acids. The elevated level of free fatty acids in women with preeclampsia may contribute to increased vascular cell adhesion molecule expression in vivo.
Adult↗