Determination of fetal rhesus d status by maternal plasma DNA analysis.

In this study, we assessed the feasibility of fetal RhD genotyping by analysis of cell-free fetal DNA(cffDNA) extracted from plasma samples of Rhesus (Rh) D-negative pregnant women by using real-time polymerase chain reaction (PCR). Fetal genotyping was performed on 30 RhD-negative women between 9 and 39 weeks of gestation who were referred to us for invasive testing [amniocentesis/chorionic villi sampling (CVS)]. The fetal RHD genotype was determined based on real-time PCR method. Exons 7 and 10 of the RHD and SRY genes were targeted. Among the pregnant women, 12 were carrying male and 17 were carrying female fetuses. Out of 29 pregnant women, 21 had RhD-positive and nine had RhD-negative fetuses. One sample (case 12, whose blood group was found to be AB Rh [+]) was excluded due to controversial results from repeated serological analyses. All prenatal results were in concordance with postnatal RhD status and fetal sex without false- positive or -negative results. Performing real-time PCR on cffDNA showed accurate, efficient and reliable results, allowing rapid and high throughput non invasive determination of fetal sex and RhD status in clinical samples.

Vaginal fluid pH, cervicovaginitis and cervical length in pregnancy.

AIM: The purpose of this prospective study was to determine the possible association among vaginal fluid pH, cervicovaginitis and cervical length in singleton pregnancies at 16-22 weeks of gestation. METHODS: A total of 240 asymptomatic singleton pregnancies at 16-22 weeks of gestation were included to the study. Vaginal fluid pH was determined using pH paper in a sterile speculum examination, and cervical length was examined by transvaginal ultrasonographic measurement. Vaginitis was diagnosed by pH determination and wet mount smear; cervicitis was diagnosed by cervical examination. Patients were followed to delivery and hospital records were reviewed to extract obstetric information. Preterm delivery was defined as delivery at or prior to 36 weeks of gestation. Abnormal pH was defined as a pH of > 5.0. Patients with cervicovaginitis (n = 72) were compared with those without any trace of infection (n = 60). RESULTS: The mean gestational age was 20.3 +/- 1.4. We found an significant association among cervicovaginitis, cervical length and vaginal pH. There was a significant correlation between an elevated vaginal pH (> 5.0) and a shortened cervical length (r = -0.59, p < 0.001). Vaginal fluid pH > 5.0 was associated with increased risk of preterm delivery (OR 4.3, 95% CI 2.0, 9.3; p = 0.001) as well as delivering an infant of less than 2,500 g (OR 4.0, 95% CI 1.4, 11.0; p = 0.009). CONCLUSIONS: Elevated vaginal fluid pH in women at 16-22 weeks of gestation seems to be associated with a decreased cervical length and increased risk of preterm delivery.

MMAC tumor supressor gene expression in ovarian endometriosis and ovarian adenocarcinoma.

OBJECTIVE: The aim of this study was to investigate the role of MMAC1 protein in the relationship between ovarian endometriosis and clear cell and endometrioid-type ovarian adenocarcinomas. METHODS: A total of 63 subjects who underwent surgery for a pelvic tumoral mass, 30 of whom were diagnosed with grade 1 to 3 ovarian adenocarcinoma and 33 of whom were diagnosed with grade 1 to 4 endometriosis during histopathological examination were included in this study. The mean age for subjects with ovarian endometrioid type adenocarcinoma was 51.8 +/- 12.4, whereas the mean age for subjects with ovarian clear cell type adenocarcinoma was 59.5 +/- 13.7. Ovarian carcinomas were graded in accordance with the FIGO 1989 grading system. The mean age for subjects with endometriosis was 37 +/- 11.9. New sections were obtained from paraffin blocks in the archives of Ege University, School of Medicine, Department of Pathology onto lysinated slides and immunohistochemical staining by using mouse monoclonal antibody (MMAC1, 28H6 clone, Novocastra, UK) as MMAC antibody was applied in order to determine MMAC1 protein. Brown staining on the nucleus was considered as positive immunoreactivity. Immunoreactive staining was evaluated as percentage staining over the whole preparative. RESULTS: Of the 63 subjects included in the immunohistochemical study, ovarian endometrioid adenocarcinoma was identified in 18 subjects, while 12 subjects were diagnosed with ovarian clear cell adenocarcinoma and 33 subjects with ovarian endometriosis. No significant relationships were observed between age and MMAC immune staining in the ovarian endometrioid adenocarcinoma (r = -0.41, p = 0.08) and ovarian endometriosis (r = 0.12, p = 0.50) groups, whereas a significant relationship was observed in the ovarian clear cell adenocarcinoma group (r = 0.631, p = 0.02). No significant relationships were observed between CA125 levels and MMAC immune staining in the ovarian endometrioide adenocarcinoma (r = 0.056, p = 0.82), ovarian endometriosis (r = 0.21, p = 0.36) and ovarian clear cell adenocarcinoma (r = 0.363, p = 0.24) groups. No correlations were observed between endometriosis stages and the MMAC immune staining (r = -0.17, p = 0.92). There was no correlation between mean diameter of endometrioma and MMAC immune staining (r = -0.230, p = 198). Mean endometrioma diameter was 5.7 +/- 3.5 (1-15.5). No correlations were detected between MMAC immune staining and ovarian endometrioide adenocarcinoma or ovarian clear cell adenocarcinoma stage (r = -0.22, p = 0.37; r = 0.44, p = 0.14, respectively). No significant relationships with respect to MMAC immune staining were detected between the endometriosis and ovarian clear cell adenocarcinoma groups (p = 0.05) and between the ovarian clear cell adenocarcinoma and ovarian endometrioid adenocarcinoma groups (p = 0.27). A significant relationship with respect to MMAC immune staining was observed between ovarian endometrioide adenocarcinoma and endometriosis groups (p = 0.001). CONCLUSION: Immunohistochemical determination of MMAC defective protein expressions could be considered for utilization as a new, simple and useful technique in determination of endometriosis patients with increased risk of malignant transformation, patients where early surgical treatment would be necessary and patients that should be subjected to follow-up controls with a higher frequency.

Evaluation of VEGF in placental bed biopsies from preeclamptic women by immunohistochemistry.

OBJECTIVE: The aim of the study was to determine VEGF protein with immunohistochemical staining in placental bed biopsies of preeclamptic pregnancies in comparison to normal controls. DESIGN: Prospective cohort study. METHODS: The placental bed biopsies were obtained from 12 patients with preeclapmsia and ten patients for a control group at the time of cesarean delivery. Tissue samples of the placental bed were examined for VEGF protein distribution with avidin-biotin-peroxidase immunohistochemistry. Two blinded histopathologists were asked to score each sample for the intensity of staining and the number of cells stained in a randomly selected HPF of each sample. The resulting "H-score" was computed as a product of intensity and percent of cells stained. RESULTS: VEGF expression was significantly lower in both the myometrium and stroma of the preeclamptic group compared to the control group (77.2 +/- 25.4 vs 134 +/- 44.3, p = 0.007; 194.1 +/- 20.7 vs 170.2 +/- 17, p = 0.017, respectively). CONCLUSION: VEGF expression is significantly lower in placental bed biopsies of preeclamptic pregnancies.

Trends in cytogenetic prenatal diagnosis in a reference hospital in Izmir/Turkey: a comparative study for four years.

The aim of the study was to investigate the major changes in the indications, culture success and abnormality rate for conventional cytogenetic prenatal diagnosis on amniotic fluid samples between the period of January 1998 and December 2001 in Izmir. The cytogenetic laboratory provides a prenatal service to obstetrics-gynecology departments of different hospitals in Izmir. A limited number of patients (6-8 per week) is randomly accepted for prenatal cytogenetic study. Over the 4 years period 1190 prenatal cytogenetic tests were performed in our center. The most common indication was advanced maternal age for each year. However its rate has increased significantly within the years (35.68% in 1998, 61.38% in 2001), while the rate of both triple test and ultrasound scanning indications decreased. Culture success rates have improved (97.97% in 1998, 99.74% in 2001). Comparing the first two years to the last two years the rate of abnormal cytogenetic results decreased significantly (3.83% in 1998-99, 2.48% in 2000-01). The major reason for this decrease is probably related to the changes in indications throughout the years.

The effect of medroxyprogesterone acetate and heparin in the prevention of postsurgical adhesion formation in the rat uterine model.

OBJECTIVE: To evaluate the effects of depot medroxyprogesterone acetate and heparin in preventing postsurgical adhesion formation in the rat model. METHODS: A hundred and five female Wistar rats were divided into 7 groups. Groups 1 and 2 were injected with intramuscular 15 mg medroxyprogesterone acetate 3 weeks before surgery and at the end of laparotomy. Groups 3 and 4 were given 15 mg medroxyprogesterone acetate by intramuscular injection, 3 weeks before surgery. An equal volume of intramuscular sterile saline was injected to control groups, 3 weeks before and at the end of surgery. Before abdominal closure, 2 ml of Ringer's lactate was instilled into the peritoneal cavity of all rats, except group 7. Groups 1, 4, and 5 were given 2 ml of intraperitoneal Ringer's lactate containing 500 U heparin/ml. A standardized surgical injury was performed in all rats. Two weeks after surgery, the adhesions were scored on a scale of 0 to 3 according to their thickness-tenacity and vascularity. Kruskall-Wallis and Mann-Whitney U statistical test were used. RESULTS: The preoperative and postoperative administration of medroxyprogesterone acetate resulted in the least number of and the least severe adhesions, when compared with single dose medroxyprogesterone acetate treated rats and controls (p < 0.05). However, the combination of medroxyprogesterone acetate and intraperitoneal heparin did not enhance the adhesion reducing capacity of medroxyprogesterone acetate. CONCLUSIONS: Concurrent preoperative and postoperative administration of medroxyprogesterone acetate results in the most significant reduction of postsurgical adhesions. The combination treatment of medroxyprogesterone acetate and heparin does not show any additional effect in the reduction of adhesion formation, when compared with medroxyprogesterone acetate treatment alone.

The comparison of uterine artery Doppler velocimetry with the histopathology of the placental bed.

We determined the relationship between the histopathological findings of the placental bed and Doppler flow measurements of the uterine artery in women with preeclampsia and fetal growth retardation. Doppler velocimetry in the uterine artery was evaluated in 17 pregnant women with preeclampsia, 15 of whom had fetal growth retardation, and 20 normal pregnant women, within 14 days of Caesarean delivery and placental bed biopsy. The placental bed biopsies were evaluated in terms of trophoblast migration into the myometrium and physiological changes of the spiral arteries. The results were compared with Doppler velocimetry values. Trophoblast migration and physiological changes were not detected in 10 (59%) cases with preeclampsia and in 4 (20%) with normal pregnancies (p<0.05). In the preeclamptic group, 9 of 15 cases that were complicated with intrauterine growth retardation had no trophoblastic migration into the myometrium. The mean systolic/diastolic ratio, resistance index and pulsatility index of the uterine artery in women with preeclampsia and fetal growth retardation was significantly higher than women with normal pregnancies (p<0.01). The mean resistance index of the uterine artery in the impaired migration group was significantly higher than the migration group (p=0.02). The incidence of impaired trophoblast migration was significantly higher in the group with a high systolic/diastolic ratio (above 2.5) and resistance index (above 0.58) than cases with low systolic/diastolic ratio and resistance index (72%, 23% respectively, p<0.05). The incidence of early diastolic notch in the impaired trophoblast migration group was significantly higher than the migration group (57% versus 13%, p<0.01). Our study supports the hypothesis that high uterine artery flow resistance is related to the reduced trophoblast migration into the myometrium and inadequate physiological changes in the spiral arteries in women with intrauterine growth retardation and preeclampsia.

Prenatal sonographic detection of nasopharyngeal teratoma.

We present the case of a 34-year-old pregnant woman who had an elevated maternal serum alpha-fetoprotein level and sonographic findings of a semisolid mass protruding from the fetus's oral cavity. The large, heterogeneous mass filled the oropharynx and nasopharynx. Abnormal Doppler waveforms were detected in the umbilical artery of the fetus, who died in utero. Postmortem examination revealed a nasopharyngeal teratoma.

Impaired antioxidant activity in women with pre-eclampsia.

OBJECTIVE: To investigate antioxidant activity of sera and the plasma blood levels of two potent antioxidant in women with pre-eclampsia and normotensive pregnancies. STUDY DESIGN: The antioxidant activity of sera and the blood levels of ascorbic acid and alpha-tocopherol were assayed in women with normal pregnancies (n = 33), mild pre-eclampsia (n = 8), and severe pre-eclampsia (n = 16) between 20 and 40 weeks' gestation. Ascorbic acid and alpha-tocopherol concentrations were analyzed by high-performance liquid chromatography. Antioxidant activity of sera was measured as the percent inhibition of spontaneous autoxidation of a standard brain homogenate. RESULTS: Plasma levels of ascorbic acid in women with mild and severe pre-eclampsia were significantly lower than normal pregnancies (P < 0.05). Sera alpha-tocopherol levels were significantly decreased only in severe pre-eclampsia (P < 0.05). Sera antioxidant activity were significantly decreased in mild (73%) and severe (51%) pre-eclampsia compared with normal (86%) pregnancies (P = 0.02, P = 0.000, respectively). CONCLUSIONS: In women with pre-eclampsia, sera antioxidant activity and antioxidant level of plasma are decreased when compared with normotensive pregnancies. Impaired antioxidant activity and the reduction of antioxidant levels which increase the level of lipid peroxidation products may cause peroxidative damage of vascular endothelium and result in clinical symptoms of pre-eclampsia.

Comparison of epidural and general anaesthesia for elective caesarean delivery according to the effects of apgar scores and acid-base status.

The objective of this study was to determine the effects of lumbar epidural anaesthesia on the Apgar score and acid-base status of the newborn. Umbilical artery blood gases were obtained in 85 singleton, term, uncomplicated pregnancies delivered by elective Caesarean section. The umbilical artery blood pH, PaCO2, PaO2 and HCO3 values and Apgar scores (1 and 5 minutes) were compared between lumbar epidural and general anaesthesia groups. General anaesthesia was used in 45 (52.9%) women and lumbar epidural anaesthesia in 40 (47.1%). Only 2 of the newborns exposed to epidural anaesthesia had umbilical artery blood pH values 7.19 or less. The mean umbilical artery blood pH was found to be significantly lower in the newborns exposed to lumbar epidural anaesthesia (p = 0.011). None of the newborns in the 2 groups were severely depressed (Apgar scores less than 4). The mean umbilical artery blood PaCO2, PaO2 and HCO3 values did not show any significant difference between the groups. In conclusion, lumbar epidural anaesthesia is associated with lower umbilical artery blood pH values, occasionally with severe fetal acidaemia.