Significance of serum c-erbB-2 oncoprotein, insulin-like growth factor-1 and vascular endothelial growth factor levels in ovarian cancer.

OBJECTIVES: Our aim was to determine the predictive values of serum levels of several growth factors in ovarian cancer, including soluble c-erbB-2 oncoprotein, insulin-like growth factor-1 (IGF-1) and vascular endothelial growth factor (VEGF). BACKGROUND: Previous studies have shown that growth factors play an important role in carcinogenesis. METHODS: Two groups were established. One of them was the malignant group which included 41 patients with ovarian carcinoma and the other was the control group that was made up of 28 healthy volunteers. Preoperative serum samples were obtained from the patients, and c-erbB-2, IGF-1 and VEGF levels were measured in these samples using ELISA. Serum CA-125 levels were also determined, by chemiluminescent microparticle immunoassay. RESULTS: VEGF levels of the malignant group were significantly higher than those of the control group (p < 0.01). CA-125 levels were also significantly higher than the in control group (p < 0.001). Area under the ROC curve (AUC) was 0.982 for CA-125, 0.780 for VEGF, 0.603 for c-erbB-2, and 0.467 for IGF-1 in differentiating cancers from controls. CONCLUSION: Serum VEGF levels might be a predictor for diagnosis in ovarian cancer patients, while serum c-erbB-2 and IGF-1 levels do not have a clinical significance in terms of ovarian cancer (Tab. 1, Fig. 1, Ref. 46).

Lobular carcinoma of the breast metastasizing to leiomyoma in a patient under letrozole treatment.

Metastasis of extragenital neoplasms to the uterus is extremely rare. Lobular breast cancers metastasize to the uterus more than ductal carcinomas, but they metastasize as tiny nodules that can be missed with the standard diagnostic workup. Uterine involvement by a metastatic tumor is usually a manifestation of end-stage disease; patients are reported to die within weeks to months. Therefore surgery is not recommended. Here we report a case of lobular breast cancer metastasizing to a leiomyoma in a patient using letrozole. Our patient was submitted to surgery because the leiomyoma had grown to the level of the xiphoid process. She is alive one year after the operation. In conclusion growth of leiomyomas under aromatase inhibitors should be considered as a sign of metastases and surgery can be planned in selected cases.

Detection of fetal RHD pseudogene (RHDΨ) and hybrid RHD-CE-Ds from RHD-negative pregnant women with a free DNA fetal kit.

Hemolytic disease of the newborn is a clinical condition in which maternal and paternal Rh blood group antigens are incompatible and the mother is negative for the antigen whereas the father is positive. Analysis of fetal cells recovered from maternal plasma can provide a highly sensitive prenatal diagnosis. The fetal RHD gene in plasma DNA is detected by real-time PCR amplification of two different segments of the RHD gene (exons 7 and 10). Each amplicon is revealed with specific probes. We examined 40 female blood samples to verify the specificity of RHD exons (7 and 10) amplified by real-time PCR. Thirty fetuses were predicted to be RHD-positive based on analysis of plasma DNA. Seven fetuses were predicted to be RHD-negative. One fetus was negative for RHD on exon 10, and positive for RHD on exon 7 (early gestation age); two fetuses were RHD-negative on exon 7, and RHD-positive on exon 10 (RHD-CE-D(s) or RHDΨ), indicative of a maternal RHD allele. We conclude that it is necessary to analyze at least two exon regions in the RHD gene.

Serum microRNA expression in pregnancies with preeclampsia.

Preeclampsia continues to be a mortal disease of pregnant women throughout the world. Recently, geneticists, allied with obstetricians, have opened new frontiers. MicroRNAs (miRNAs) are members of a class of small, noncoding RNA molecules. They are critical posttranscriptional regulators of gene expression. We extracted circulating miRNA from maternal plasma and quantified mir-152 and mir-210. We found up-regulated miR-210 levels as well as down-regulated mir-152 levels in preeclampsia patients.We propose that detection of increased mir-210 levels in maternal serum could be used to improve prediction methods for noninvasive prenatal diagnosis of preeclampsia.

Vascular endothelial growth factor in adnexal masses.

OBJECTIVES: To determine cyst fluid and serum vascular endothelial growth factor (VEGF) concentrations in patients with ovarian masses and to investigate the efficiency of this modulator in the clinical management of cystic pelvic masses. METHODS: Needle puncture for cyst fluid aspiration were performed on 88 cystic ovarian masses intraoperatively. Forty-five patients with benign and 43 patients with malignant ovarian pathology were analyzed for cyst fluid and serum VEGF concentrations. Both cystic fluid and serum VEGF concentration were determined by enzyme-linked immunosorbent assay (ELISA). RESULTS: Cyst fluid VEGF levels of malignant cysts (40.65+/-17.69 ng/ml) were significantly higher than those of benign cysts (12.53+/-6.13 ng/ml; P<0.001). Similarly, higher serum VEGF concentrations were found in patients with malignant disease (0.72+/-0.17 ng/ml) compared with benign cysts (0.33+/-0.11 ng/ml; P<0.001). A statistically significant correlation was observed between cyst fluid and serum VEGF levels in both malignant and benign cysts. For serum VEGF, at a cut-off value of 0.41 ng/ml; sensitivity, specificity, PPV, and NPV were 95%, 78%, 80% and 95%, respectively. No significant correlation between cyst fluid VEGF concentration and tumor stage or grade could be found. CONCLUSIONS: Significantly higher concentrations of VEGF are present in cyst fluid and serum of patients with malignant ovarian cysts compared with benign ovarian ones. There is no relation between VEGF and tumor stage or grade.

Prenatal prediction of childhood-onset spinal muscular atrophy (SMA) in Turkish families.

Childhood-onset spinal muscular atrophy (SMA) is one of the most common neurodegenerative genetic disorders. SMN1 is the SMA-determining gene deleted or mutated in the majority of SMA cases. There is no effective cure or treatment for this disease yet. Thus, the availability of prenatal testing is important. Here we report prenatal prediction for 68 fetuses in 63 Turkish SMA families using direct deletion analysis of the SMN1 gene by restriction digestion. The genotype of the index case was known in 40 families (Group A) but unknown in the remaining 23 families (Group B). A total of ten fetuses were predicted to be affected. Eight of these fetuses were derived from Group A and two of these fetuses were from Group B families. Two fetuses from the same family in Group A had the SMNhyb1 gene in addition to homozygous deletion of the NAIP gene. One fetus from Group A was homozygously deleted for only exon 8 of the SMN2 gene, and further analysis showed the presence of both the SMN1 and SMNhyb1 genes but not the SMN2 gene. In addition, one carrier with a homozygous deletion of only exon 8 of the SMN1 gene was detected to have a SMNhyb2 gene, which was also found in the fetus. To our knowledge, these are the first prenatal cases with SMNhyb genes. Follow-up studies demonstrated that the prenatal predictions and the phenotype of the fetuses correlated well in 33 type I pregnancies demonstrating that a careful molecular analysis of the SMN genes is very useful in predicting the phenotype of the fetus in families at risk for SMA.

Apoptosis in the placenta of pregnancies complicated with IUGR.

OBJECTIVE: In this study we have investigated the presence of apoptosis in the placental tissue of pregnancies complicated with intra-uterine growth restriction (IUGR). METHOD: Placental samples were obtained from 22 normal third trimester pregnancies and 20 pregnancies complicated with IUGR. The criteria for fetal growth impairment were clinical evidence of sub-optimal growth, ultrasonographic demonstration of deviation from normal percentiles of growth and birth weight under 10th percentile. Terminal deoxynucleotidyl transferase mediated deoxyuridine triphosphate nick end labelling (TUNEL) staining was used to demonstrate the apoptotic cells in all samples. Student-t, Mann-Withney U-test, Fisher exact test and Spearman correlation were used for statistical analysis. RESULTS: We detected apoptosis in 10 placentas in the study group vs. none in the control group. Placentas from pregnancies complicated with IUGR demonstrated 0.12% (0.1%-0.4%) apoptotic cells. The rate of apoptotic cells in the placenta was significantly higher in pregnancies complicated with IUGR than normal uncomplicated pregnancy (P=0.0019). Apoptosis were more abundant in the trophoblasts, especially cytotrophoblasts, in the placenta. We could not find a correlation between the apoptosis in the placenta of pregnancies complicated with IUGR and birth weight, multi-parity, gestational age, birth weight percentile and mode of delivery (C/S vs. vaginal delivery). CONCLUSION: We believe that the increased number of apoptosis in the placenta of pregnancies complicated with IUGR may have an important compensatory role to transmit nutrition and gas exchange easily to the fetus.

Perinatal and maternal outcomes of fetal macrosomia.

OBJECTIVE: To determine the perinatal and maternal outcome of the macrosomic infants. STUDY DESIGN: A case-control, retrospective study is performed in the Department of Gynecology and Obstetrics, Istanbul University Cerrahpasa Medical Faculty, between 1988-1992. The maternal and neonatal records of infants with birthweight of at least 4000g (n=1000) were reviewed. Another 1000 cases amongst the newborns delivered in the same period between 2500 and 3999g formed the control group. The obstetrical outcome variables of the groups including mode of delivery and the incidence of maternal and perinatal complications were compared. RESULTS: A total of 16,112 deliveries occurred during the study period. The rate of macrosomic deliveries was 6.21% and the rate of the deliveries (4500g or heavier) was 1.04%. The mean birthweight of the study group was 4272+/-239 and 3277+/-316g of the control group (P<0.001). While the cesarean section rate was 28.8% for the study group and it was 16.6% for the control group (P<0.001). In the study group, 17 cases of brachial plexus palsy (2.4%), 16 cases of clavicular fracture (2.3%) and one case of humeral fracture were observed (P<0.001). The rate of perinatal mortality was 0.8% in the study group. No perinatal mortality was recorded in the control group. There were 14 cases (1.4%)of asphyxia related to delivery in the study group (P<0.01). The rate of maternal complications, were significantly higher in the study group (P<0.01). CONCLUSION: The macrosomic infants are in increased risk for birth trauma and asphyxia. The risk of birth trauma for the infants weighing 4500g or more is even greater.

High levels of fetal erythroblasts and fetal extracellular DNA in the peripheral blood of a pregnant woman with idiopathic polyhydramnios: case report.

Abnormal amniotic fluid volume can be associated with increased maternal risk as well as perinatal morbidity and mortality. Polyhydramnios is often indicative of fetal, placental or maternal problems. In a large proportion of patients the aetiology of the disorder is unclear. Here we report on a case in which numerous fetal erythroblasts and large quantities of extracellular fetal DNA were found in the peripheral blood of a pregnant woman with idiopathic polyhydramnios bearing a male fetus. Following enrichment of erythroblasts by magnetic separation (MACS) and anti-CD71 antibodies, approximately 45-fold more erythroblasts were determined per ml peripheral maternal blood than in matched controls (231 versus 5). Single cell multiplex polymerase chain reaction (PCR) of individually micromanipulated erythroblasts showed that approximately 122 of these were of fetal origin. The concentration of extracellular fetal circulatory DNA in maternal plasma was determined by real-time quantitative PCR and shown to be almost double that of the control group (749.2 versus 404 fetal genome equivalents per ml maternal plasma). It can be speculated that the increased intrauterine pressure in polyhydramnios leads to an enhanced influx of fetal cells and free extracellular fetal DNA into the maternal circulation. This hypothesis will have to be tested with further cases.

Evaluation of the fetal kidney maturation by assessment of amniotic fluid alpha-1 microglobulin levels.

OBJECTIVE: To evaluate the fetal renal maturation by assessment of amniotic fluid microproteins and to show these proteins originate from fetal urine. STUDY DESIGN: Amniotic fluid proteins (total protein, albumin, high molecular weight protein-HMWP, low molecular weight protein-LMWP, alpha(1)-microglobulin and beta(2)-microglobulin) were determined in 39 pregnant women at delivery and by amniocentesis in 30 pregnant women. These values were compared with first urine values of neonates with the same gestational age. RESULTS: Albumin was the largest protein component in the amniotic fluid. LMWP showed an increase in the amniotic fluid until the end of the second trimester; and as pregnancy advanced a progressive decrease occurred in parallel to fetal renal maturation. After 26 weeks' gestation, a strong correlation was identified between LMWP levels and alpha(1)-microglobulin, and between LMWP and beta(2)-microglobulin. No significant difference was detected between LMWP levels in the first urine of the neonates and in amniotic fluids. CONCLUSION: Microproteins in the fetal urine are of fetal origin. Fetal renal maturation can be evaluated by measuring microproteins in the amniotic fluid. Fetal renal maturation is best reflected by alpha(1)-microglobulin.

A strong prognostic variable in endometrial carcinoma: flow cytometric S-phase fraction.

BACKGROUND: The purpose of this study was to determine the role of flow cytometric S-phase fraction as a prognostic factor in patients with endometrial adenocarcinoma. METHODS: The study included 80 patients with endometrial adenocarcinoma of endometrioid type who were followed regularly between 1984-1995 in the Department of Obstetrics and Gynecology at Cerrahpasa Faculty of Medicine in Istanbul, Turkey. The method employed for the flow cytometric analysis was modified from Hedley et al. RESULTS: The S-phase fraction was identified as the most significant variable associated with death from endometrial carcinoma of endometrioid type by the Cox proportional hazards model. The risk of death was significantly higher in patients with S-phase values greater than 20%. Aneuploidy and DNA indexes were also significant prognostic variables. CONCLUSIONS: The S-phase fraction is considered to be a significant prognostic variable in identifying those patients with endometrial carcinoma who have a poor prognosis. The authors believe that S-phase fraction distinguishes those patients who may benefit from additional treatment approaches.

DNA ploidy of ectopic pregnancy and first trimester spontaneous abortion investigated by flow cytometry.

BACKGROUND: To compare the success rate of DNA flow cytometry in determining the DNA ploidy status in ectopic pregnancy and first trimester spontaneous abortion. METHODS: Thirteen women with ectopic pregnancy (Group I) and 17 women with first trimester spontaneous abortion (Group II) were included into this study. DNA flow cytometric analysis was performed on all specimens. Aneuploidy was classified according to DNA index. The first trimester spontaneous abortions were also karyotyped after long-term culture of chronic villi. Student-t test and Fisher's exact test were used in statistical comparisons. RESULTS: DNA aneuploidy was found in five women with ectopic pregnancy (38.5%) versus in 12 women with first trimester spontaneous abortion (70.6%), and it was comparable. A triploidy and a tetraploidy were detected in group I. Six tubal ectopic pregnancies were unruptured at laparatomy and four of them had aneuploid DNA content. CONCLUSIONS: We believed that DNA flow cytometry was successful in determining the ploidy status of ectopic pregnancy and first trimester spontaneous abortion. In addition, it was interesting that ectopic pregnancies with aneuploid DNA content tended to be unruptured. However, this suggestion needs to be confirmed by further studies with larger numbers of cases.

The prognostic significance of maternal serum CA125 measurement in threatened abortion.

The prognostic predictive value of maternal serum CA125 measurement was investigated in 25 cases of threatened abortion. The women were non-smoker, had a ultrasonographically verified viable single fetus, and the gestational ages ranged from 7 to 12 weeks. Twenty-five healty pregnant women, with the same characteristics were used as the control group. The overall abortion rate was found to be 20% (5/25) in the study group. In serial measurements the mean serum CA125 level of the patients with an unfavorable pregnancy outcome was significantly higher than that of the patients with a favorable outcome. When the cut-off level of maternal serum CA125 was taken as > 65 U/ml in the first and > 60 U/ml in the second measurements of the study group, the risk of termination of the pregnancy by spontaneous abortion was 83.3% in the patients with elevated serum CA125 levels. No statistically significant difference was observed with respect to the duration of vaginal bleeding between the aborters and the patients with a favorable outcome. Nevertheless, when vaginal bleeding had been present for 3 days or more and there was high maternal serum CA125 activity, the abortion risk was found to be 100% (3/3). These findings suggest that the maternal serum CA125 measurement in threatened abortion can be useful to determine the extent of decidual destruction which is directly related to the outcome of pregnancy.

[Electron microscopic study of the amniotic epithelium in polyhydramnios and oligohydramnios]. / Die elektronenmikroskopische Untersuchung des Amnionepithels bei Polyhydramnion und Oligohydramnion.

Totally 25 cases without any fetal anomaly, 10 with polyhydramnios, 10 with oligohydramnios and 5 with normal volume of amniotic fluid were taken into consideration. Their amnion covering the placenta and umbilical cord were examined under electron microscope. In the group with polyhydramnios the microvilli facing the amniotic cavity were denser in certain regions. The intercellular space was widened and the terminal bars were opened. Within the cells the number of vesicles were increased and there were large cysternas within these vesicles. It was postulated that the large cysternas found in the basal and apical parts of the cell were composed of macropinocytotic vesicles of the basal membrane. In the group with oligohydramnios the microvilli on the apical side were diminished. The intercellular space of the lateral side was narrowed. The electron density of the basal lamina was increased. The cellular structures were apparently reduced having just a few vesicles and lipid granules. Both in polyhydramnios and oligohydramnios the amniotic epithelium cells covering the placenta and umbilical cord are responsible for the transfer of the fluid into the amniotic cavity. Possibly they control the amount of fluid by reducing or increasing its passage.

Selective fetocide by direct intracardiac injection of isotonic potassium chloride.

In multiple pregnancies with one abnormal fetus the active options are termination of pregnancy or selective fetocide. The different methods of selective fetocide are discussed and the intracardiac instillation of potassium chloride is described in a case report. Coagulation disturbances were not observed during follow-up.