Clinical application of fetal sex determination using cell-free fetal DNA in pregnant carriers of X-linked genetic disorders.

As the first step in prenatal diagnosis of X-linked genetic disorders, chorionic villus sampling (CVS) for fetal sex determination is generally performed at 11-13 weeks of gestation. However, as the procedure-related miscarriage rate of CVS is 0.5-1.0%, non-invasive methods such as PCR of cell-free fetal DNA (cff-DNA) in maternal plasma are preferable. Here, we determined fetal sex at 9-12 weeks of gestation using PCR of cff-DNA in three pregnant carriers of Duchenne muscular dystrophy. The fetal sex was accurately determined in all three cases, as confirmed by ultrasound and amniocentesis at 16 weeks (for the two female fetuses) and CVS at 12 weeks (for the one male fetus). This procedure could avoid unnecessary CVS in female fetuses.

Effect of placenta previa on blood loss in second-trimester abortion by labor induction using gemeprost.

OBJECTIVES: The study was conducted to determine whether placenta previa increases bleeding during gemeprost-induced termination of second-trimester pregnancy. METHODS: We carried out a retrospective study of 158 second-trimester terminations between 12 and 21 weeks' gestation. We compared the intraoperative blood loss among three groups: women without placenta previa undergoing gemeprost termination, women with placenta previa undergoing gemeprost termination and women with placenta previa undergoing dilatation and evacuation (D&E). RESULTS: Eleven of 158 women (7.0%) had placenta previa; four underwent D&E and seven had gemeprost termination. There was no statistical difference in mean intraoperative blood loss among the three groups, although one of the seven women with placenta previa who underwent gemeprost termination developed serious bleeding requiring blood transfusion. CONCLUSION: The use of gemeprost for second-trimester pregnancy termination in women with placenta previa seems to be relatively safe and does not increase intraoperative blood loss in the majority of cases.

Tumors Sharply Increased after Ceasing Pazopanib Therapy for a Patient with Advanced Uterine Leiomyosarcoma: Experience of Tumor Flare.

Pazopanib has activity in patients with soft-tissue sarcoma. We report an advanced uterine leiomyosarcoma case that suddenly worsened after cessation of pazopanib therapy. A 47-year-old woman had a primary uterine leiomyosarcoma tumor and multiple lung metastases, which progressed during her initial treatment. In subsequent treatment with pazopanib for 3 months, the sum of her tumor diameters after cessation sharply increased for two weeks. Symptoms such as dyspnea suddenly worsened also. She died of the disease one month after cessation of pazopanib therapy. Given the poor prognosis of recurrent uterine leiomyosarcoma and the rapid tumor enlargement after ending pazopanib therapy, control of this disease is especially important. Therefore, the decision to discontinue pazopanib therapy requires careful consideration.

Congenital diaphragmatic hernia: an evaluation of the prognostic value of the lung-to-head ratio.

PURPOSE: A retrospective analysis of the prognostic significance of the lung-to-head ratio (LHR) on the outcome of fetuses with left-sided congenital diaphragmatic hernia (CDH). METHODS: A total of 12 fetuses with isolated left CDH without any chromosomal abnormalities were included in this study. Twelve LHR measurements could retrospectively be calculated from the last available ultrasonographic recordings before birth. The relationship between the LHR and fetal outcome and gestational age was investigated. The cutoff levels as previously published were applied to determine their predictive value in this population. The association between other prenatal predictive variables and the fetal outcome was also determined. Survival was defined as being discharged from the hospital. RESULTS: The overall survival rate was 75%, and a statistically significant difference was observed between the mean LHR of the survivors and the mean LHR of the nonsurvivors (1.81 vs. 0.43), whereas the mean gestational age of these two groups did not differ. The LHR was not gestational age dependent with regard to its ability to predict the fetal outcome. The cutoff levels LHR <1, 1-1.4, and >1.4 showed good applicability for predicting fetal outcome within the present study population, with 100% survival if LHR >1.4 and 75% mortality if LHR <1. The intrathoracic position of the stomach, mediastinal shift, and polyhydramnios as individual variables and early diagnosis (<25 weeks gestation) all turned out to be poor sonographic predictors of fetal outcome. CONCLUSION: The LHR proved to be a good predictor of fetal outcome, independent of gestational age at the time of measurement. To further substantiate our observations, a prospective multicenter study is warranted.

A monozygotic twin pregnancy discordant for acardia and X-inactivation pattern.

Monochorionic, diamniotic twins discordant for acardia are reported. A 775 g acardius was dead, generally hydropic and severely malformed, while the other twin with a weight of 1314 g was phenotypically normal. Both twins had a normal 46 XX karyotype. Genotyping at 24 autosomal polymorphic loci revealed that the twins were identical (monozygotic) and ruled out uniparental disomy for chromosome 2. X-inactivation studies showed a skewed pattern in the acardiac twin and a normal random pattern in the co-twin, suggesting that the underdevelopment of the acardiac twin may have reflected the small number of original cells that were separated from the inner cell mass.

Microarray comparative genomic hybridization (CGH)-based prenatal diagnosis for chromosome abnormalities using cell-free fetal DNA in amniotic fluid.

Cell-free fetal DNA (cffDNA) in the supernatant of amniotic fluid, which is usually discarded, can be used as a sample for prenatal diagnosis. For rapid prenatal diagnosis of frequent chromosome abnormalities, for example trisomies 13, 18, and 21, and monosomy X, using cffDNA, we have developed a targeted microarray-based comparative genomic hybridization (CGH) panel on which BAC clones from chromosomes 13, 18, 21, X, and Y were spotted. Microarray-CGH analysis was performed for a total of 13 fetuses with congenital anomalies using cffDNA from their uncultured amniotic fluid. Microarray CGH with cffDNA led to successful molecular karyotyping for 12 of 13 fetuses within 5 days. Karyotypes of the 12 fetuses (one case of trisomy 13, two of trisomy 18, two of trisomy 21, one of monosomy X, and six of normal karyotype) were later confirmed by conventional chromosome analysis using cultured amniocytes. The one fetus whose molecular-karyotype was indicated as normal by microarray CGH actually had a balanced translocation, 45,XY,der(14;21)(q10;q10). The results indicated that microarray CGH with cffDNA is a useful rapid prenatal diagnostic method at late gestation for chromosome abnormalities with copy-number changes, especially when combined with conventional karyotyping of cultured amniocytes.

Clinical outcome of infants with confined placental mosaicism and intrauterine growth restriction of unknown cause.

The purpose of this study was to know a role of confined placental mosaicism (CPM) in perinatal outcome and postnatal growth and development of infants with intrauterine growth restriction (IUGR). We selected 50 infants with IUGR (<-2.0 SD) from 3,257 deliveries in a regional medical center during the past 10-year period, and carried out cytogenetic and molecular analyses in their placenta and cord blood. Of the 50 infants, 8 had CPM (CPM group) and were composed of five single (CPM2, 7, 13, 22, and 22), one double (CPM7/13), and one quadruple trisomy (CPM2/7/15/20), and one partial monosomy [del(2)(p16)]. The origin of an extra chromosome of trisomy was maternal in six cases of CPM, paternal in one, and undetermined in one. Uniparental disomy in disomic cell lines was ruled out in all these mosaics. We also compared clinical parameters for perinatal outcome between CPM group and infants without evidence of CPM (non-CPM group), such as maternal and gestational age, birth weight, Apgar score, cord blood pH, gender, and uterine artery patterns by Doppler ultrasonography, as well as weight, height, and developmental quotient (DQ) by Denver Developmental Screening Test at age 12 months. Phenotypic abnormalities were noted in two infants with CPM and three infants of non-CPM group: One with CPM22 had ASD and hypospadias, one with CPM7/13 had Russell-Silver syndrome (RSS), and one without CPM had polydactyly, and two without CPM had RSS. All but one infant with CPM are alive at age 12 months. Among the clinical parameters, the detection rate of a notch waveform pattern of the uterine artery was significantly higher in the CPM group (P < 0.05). However, no significant difference was noted in perinatal outcome of pregnancy and in DQ at age 12 months between the two groups. Interestingly, short stature (<-2 SD) at age 12 months was more frequently seen in CPM group (7/8 infants with CPM vs. 8/15 infants without CPM), although no statistically significant difference was obtained. The information obtained will be useful for perinatal care and genetic counseling for infants with IUGR and CPM.

A monozygotic conjoined twin pregnancy discordant for laterality of cleft lip.

Conjoined twins discordant for phenotype of cleft lip are reported. One had right-sided cleft lip, while the co-twin had left-sided cleft lip. Both twins had a normal 46,XY karyotype. Genotyping at 50 polymorphic loci revealed that the twins were identical (monozygotic). Our analysis suggests that the laterality of cleft lip is more affected by the process of twinning than by genetic factors.