Polymerase-chain-reaction-based detection of fetal rhesus D and Y-chromosome-specific DNA in the whole blood of pregnant women during different trimesters of pregnancy.

OBJECTIVE: The aim of this study was to determine whether or not a noninvasive procedure utilizing maternal peripheral blood as the source of DNA and polymerase chain reaction (PCR) could be used to detect fetal rhesus D (RhD) status as well as fetal gender during different gestational stages of pregnancy. MATERIALS AND METHODS: Maternal blood samples were obtained from 54 RhD-negative pregnant women during the first trimester (6-13 weeks, n = 14), second trimester (14-26 weeks, n = 26) and third trimester (27-40 weeks, n = 14). Genomic DNA was extracted from the whole blood and analyzed by seminested and nested PCR for detection of DNA sequences corresponding to RhD (n = 54) and Y chromosome (n = 48) using RhD and Y-chromosome-specific oligonucleotide primers, respectively. The seminested/nested PCR results were compared with the RhD status and gender of the babies after delivery. RESULTS: The sensitivity and specificity of seminested PCR for detection of fetal RhD positivity in whole blood of pregnant women were 81 and 100%, respectively, while the sensitivity and specificity of nested PCR for detection of male fetuses, using Y-chromosome-specific DNA as a marker, were 96 and 91%, respectively. There were no significant differences in the PCR results with samples obtained from women at different gestational stages of pregnancy. CONCLUSION: Seminested and nested PCRs for detection of fetal RhD and gender status, respectively, by using the blood of pregnant women during different gestational stages of pregnancy, are reliable noninvasive procedures with high sensitivity and specificity.

Hormone profile of Kuwaiti women with hyperemesis gravidarum.

OBJECTIVE: To evaluate the incidence of hyperemesis gravidarum among pregnant women in Kuwait and the status of HCG, TSH, Total T4 and Free T4 in the serum of patients with hyperemesis gravidarum compared with a control group of women. METHODOLOGY: During a 6-month period all patients admitted to Maternity Hospital with features of hyperemesis gravidarum (excessive vomiting and ketonuria) were enlisted into the study. In fifty of these patients and their fifty normal controls, the status of serum total (beta)hCG, TSH, total T4 and freeT4 were evaluated with AXSYM micro particle enzyme immunoassay. RESULTS: The incidence of hyperemesis in the maternity population was 45 per 1000 deliveries. Total (beta)hCG and Total T4 and FreeT4 were significantly higher in the hyperemesis patients than in the normal controls (p<0.0001, p=0.004 and p=0.01 respectively). TSH levels were significantly lower in hyperemesis patients than in their normal controls (p<0.0001). There was a strong positive correlation between the total (beta)hCG and the gestational age (r=0.8). CONCLUSION: There is a high incidence of hyperemesis gravidarum in the Kuwaiti population. Total (beta)hCG, Total T4 and Free T4 titers were significantly higher in patients with hyperemesis gravidarum, but none of the patients showed signs of hyperthyroidism.

Effect of increased glucose load on maternal-fetal transport of alpha-aminoisobutyric acid in the perfused human placenta: in vitro study.

The role of hyperglycemia on modulation of maternal-fetal transport of amino acids in humans is little understood. Hence, we have explored the effect of increased glucose load on transport kinetics of a model non-metabolizable amino acid, alpha-aminoisobutyric acid (AIB), in the human placenta in vitro. Transport kinetics of AIB in maternal-fetal direction was studied using perfusion of isolated human placental lobules. NCTC (National Culture and Tissue Collection)-135 medium, diluted with Earle's buffered salt solution was used as the perfusate and tritiated water was used as the reference marker. Effect of increased glucose load on transport kinetics of study and reference substances was studied in normal term placentae (n=5; gestational age, 38.5 +/- 0.5 weeks) in succeeding experimental phases, after a control perfusion phase with physiological glucose concentration. AIB transport fraction (TF), relative to tritiated water TF, averaged 54.8% in control euglycemic phase while in hyperglycemic concentration phases of 27.8 and 55.6 mM, the AIB TF index averaged 42.4% and 38.2%, respectively. Analysis of variance revealed that the difference was statistically significant. Similarly, absorption rate index of the amino acid was also significantly lower in the hyperglycemic perfusion phases compared to control euglycemic phase. We conclude that hyperglycemia may play a deleterious role in limiting maternal-fetal transport of A-type amino acids in the in vivo state.

Effect of repeated doses of dexamethasone on the incidence and severity of respiratory distress syndrome in multifetal gestation between 24 and 34 weeks.

The objective of the study was to evaluate the effects of antenatal corticosteroid therapy in multiple pregnancy on the incidence and severity of respiratory distress syndrome (RDS). Twenty-two women with twin, 12 with triplet and 2 with quadruplet pregnancies and an emergency group of 20 mothers with twin pregnancies, who only had one course of dexamethasone, were compared with controls. Corticosteroid therapy was associated with a reduction in moderate to severe RDS in all groups (twins p < 0.01, triplets and quadruplets p < 0.008) and the emergency group with dexamethasone therapy (p < 0.036) when compared to their individual controls. Dexamethasone reduced the duration of stay in the neonatal intensive care unit of the emergency twin group (p < 0.01). Neonatal birth weights were significantly less in all groups treated with repeated doses in comparison to the controls.

Differential levels of interleukin 6 in maternal and cord sera and placenta in women with pre-eclampsia.

BACKGROUND: Interleukin 6 (IL-6) is a T helper 2 cytokine with a variety of properties including pro-inflammatory characteristics. It has, therefore, been implicated in the pathophysiology of abnormal pregnancies. OBJECTIVE: To investigate the association between IL-6 and pre-eclampsia by estimating the differential levels of IL-6 in maternal and cord serum and supernatant of homogenized placental tissue. METHODS: 50 primigravidae with pre-eclampsia and 50 matched normotensive primigravidae served as controls. At delivery, maternal and cord blood were collected and the serum extracted. Placental blocks were homogenized and sonicated in RPMI solution and the supernatant collected. The total protein concentration was determined and IL-6 levels assayed with an ELISA technique. RESULTS: Placental IL-6 (170 and 186 pg/mg protein) was threefold that in the maternal (64 and 58 pg/mg protein) and cord serum (63 and 72 pg/mg protein; p < 0.01). There was no significant difference in the mean IL-6 levels in maternal and cord serum or placenta in both pre-eclamptic women and normotensive controls nor in pre-eclamptic patients with babies with intra-uterine growth restriction or in pre-eclamptic patients with babies with an appropriate birth weight and in normotensive controls. CONCLUSION: There are no differences in the maternal and cord sera and placental levels of IL-6 in pre-eclamptic and normotensive women, indicating that IL-6 may not have a role in the pathophysiology of pre-eclampsia.

Detection of Y chromosome-specific DNA in the plasma and urine of pregnant women using nested polymerase chain reaction.

The present study was undertaken to evaluate a nested polymerase chain reaction (PCR) for detection of Y chromosome-specific fetal DNA in maternal plasma and urine of pregnant women during different gestational stages. DNA isolated from plasma and urine samples of 80 pregnant women (between 7 and 40 weeks' gestation) underwent amplification for Y chromosome-specific 198 bp DNA by nested PCR. The postpartum analysis of fetal gender showed that 55 women carried male and 25 female fetuses. Among the 55 women bearing male fetuses, Y chromosome-specific signals were detected in 53 (96%) plasma and 21 (38%) urine samples. Moreover, out of 25 women bearing female fetuses, 3 (12%) and 1 (4%) women had Y chromosome-specific signal in plasma and urine, respectively. Analysis of results with respect to gestational age revealed that there was no significant difference in the detection of Y chromosome-specific DNA between different trimesters in maternal plasma of women bearing male fetuses. These results showed that fetus-specific DNA was detected with high sensitivity (96%) and specificity (88%) in the maternal plasma by nested PCR, and therefore the method could be useful as a non-invasive procedure for fetal sex determination and prenatal diagnosis.

Alteration of sonographic texture of the endometrium in post-menopausal bleeding. A guide to further management.

BACKGROUND: The purpose of this study was to assess the utility of transvaginal ultrasonography in the evaluation of endometrial morphology in addition to the standard criterion of endometrial thickness for selecting patients for endometrial sampling. METHODS: Two hundred and seven consecutive cases of postmenopausal bleeding were evaluated by transvaginal ultrasound. Endometrial thickness was measured as the maximum anteroposterior thickness of the endometrium including both the anterior and posterior layers, in the sagittal long axis view. The morphology of the endometrium was studied and categorized as homogeneous, focally increased echogenecity, diffusely increased echogenecity or diffusely inhomogeneous. Patients were followed up for clinical course and endometrial histopathology. RESULTS: Textural inhomogeneity was observed in all the three cases of endometrial cancers with endometrial thickness of less than 6 mm, and, in ten out of 11 cases of a more than 6 mm thick endometrium. On the other hand the endometrial texture was homogeneous in all cases of endometrial atrophy/tissue inadequate for diagnosis, with thickness of less than 6 mm. CONCLUSION: This study adds the dimension of abnormal echogenecity of the endometrium to the currently followed criterion of endometrial thickness with a view to enhance accuracy, both for a better prediction of atrophy and a higher prediction for endometrial cancer. Expectant management can be offered to patients with a homogeneous endometrium which is 6 mm thick or less. Aggressive evaluation for a malignancy must be made if there is a focal increased echogenecity or a diffuse increased echogenecity even in a thin endometrium.

Kinetics of palmitic acid transport in insulin-dependent diabetic pregnancies: in vitro study.

BACKGROUND: The paucity of data relating to transport kinetics of free fatty acids (FFA) in pregnant diabetic women prompted the undertaking of the present study. METHODS: Transport kinetics of a model FFA, palmitic acid, have been investigated in Type I diabetic pregnancies, using in vitro perfusion of isolated placental lobules. National Cancer Tissue Culture medium diluted with Earle's buffered salt solution was used as the perfusate and control placental lobules were perfused for comparison. RESULTS: In five Type I diabetic women, the palmitic acid transport fraction (TF) averaged 5.6 +/- 0.42% of injected maternal bolus dose, representing 11.8 +/- 2.1% that of tritiated water used as reference. In control perfusions (n = 5), the palmitic acid TF represented 10.2 +/- 1.3% of tritiated water TF. Differential transport rates of palmitic acid for 10, 25, 50, 75 and 90% of efflux in fetal veins differed significantly from the corresponding values for tritiated water in both study and control series. However, palmitic acid transport rates for the various efflux fractions in the two series were not significantly different. For kinetic parameters, such as area under the curve, clearance, elimination constant, time for maximum response, absorption rate and elimination rate, the values for palmitic acid in the diabetic and control series also did not differ significantly. CONCLUSION: Transport kinetics of palmitic acid in Type I human diabetic pregnancies in in vitro conditions do not differ significantly from those observed in normal pregnancies.

Effect of glucose load on the transport kinetics of palmitic acid in the human placenta: an in vitro study.

1. A paucity of data relating to free fatty acid (FFA) transport in the human placenta in non-steady state conditions prompted us to undertake the present study. 2. The transport kinetics of palmitic acid in non-steady state conditions have been investigated in vitro using human perfused placental lobules. The effects of varying glucose concentrations on maternal-foetal transport of the FFA were also investigated to mimic the hyperglycaemic states of human diabetic pregnancies. 3. National Cancer Tissue Culture medium diluted with Earle's buffered salt solution was used as the perfusate. [14C]-Palmitic acid, along with tritiated water as a reference, was injected as a bolus into the maternal arterial perfusate and perfusate samples were collected from the venous outflow for a period of 5 min. 4. The transport fraction (TF) of palmitic acid, expressed as percentage of the injected bolus, averaged 3.45 +/- 0.15% in five perfusions, representing 9.2 +/- 1.3% of the corresponding reference marker TF. Kinetic parameters, as well as TF indices of palmitic acid expressed in relation to the reference substance, did not differ significantly between perfusions with a physiological glucose load and those with hyperglycaemic concentrations of glucose of 27.8 and 55.6 mmol/L. 5. The present study shows that hyperglycaemia per se does not significantly alter palmitic acid transport kinetics in vitro in the human perfused placental lobule.

Differential levels of T helper cytokines in preeclampsia: pregnancy, labor and puerperium.

BACKGROUND: There are suggestions that T helper 1 cytokines may be detrimental to early pregnancy and T helper 2 cytokines protective of the pregnancy. Their role in preeclamptic pregnancy, labor and puerperium, is not clear. MATERIALS AND METHODS: Twenty-eight preeclamptic women and their matched controls were evaluated, at the Departments of Obstetrics and Gynecology, Faculty of Medicine, Kuwait University and Maternity Hospital, Kuwait. Outcome measures evaluated were serum levels of TNF alpha and interleukin-4 at 12, 24, 36 weeks of gestation, advanced labor and at 1 hour and daily postpartum until they were undetectable, using ELISA technique. RESULTS: T helper cytokines showed higher serum levels in preeclampsia than normotensive pregnancy (p < 0.01, 0.01), in established labor (p < 0.05) and at 1 hour postpartum (p < 0.01 for IL-4) and p < 0.02 for TNF alpha. There was significant increase of IL-4 between 12 to 24 weeks in normal pregnancy compared to preeclampsia (p < 0.001) but not for TNF alpha. By 24 hours postpartum, IL-4 was still detectable in eight parturients compared to one patient with detectable TNF alpha (p < 0.04). Detectable IL-4 levels after 24 hours postpartum were associated with intrauterine growth retardation (p < 0.03). CONCLUSION: IL-4 has a dichotomous role in pregnancy. Normotensive pregnancy is associated with high increase in IL-4 in the first half of the pregnancy, but in the second half of pregnancy and puerperium, high levels of IL-4 are associated with preeclampsia.

Expression of the Na(+)-H+ exchanger isoform-1 and cyclooxygenases in human placentas: their implications in preeclampsia.

The sodium hydrogen exchanger isoform, NHE-1 plays an important role in electrolyte and water homeostasis. These functions are compromised in pregnancies complicated with preeclampsia. At present it is not known whether NHE-1 expression is altered during preeclampsia. In the present study the placental level of NHE-1 protein was measured using immunoblotting. Since prostaglandins regulate the secretory and absorptive functions, the levels of prostaglandin E-2 as well as the expression of cyclooxygenase-1 and -2 were also estimated. The amount of NHE-1 protein and cyclooxygenase-2 was reduced in preeclamptic placentas, whereas the level of cyclooxygenase-1 remained unaltered. In contrast, prostaglandin E-2 concentration was higher in preeclampsia. Suppression of NHE-1 might render the placenta with impaired uptake of water and electrolytes and therefore may be involved in the pathogenesis of preeclampsia. While prostaglandin E-2 may play a role in preeclampsia, these findings discount the induction of cyclooxygenase-genes for this increase.