Correlation of confined placental mosaicism with fetal intrauterine growth retardation. A case control study of placentas at delivery.

OBJECTIVE: Our purpose was to determine if the frequency of confined placental mosaicism in newborns with unexplained intrauterine growth retardation (IUGR) was higher compared with infants with appropriate growth in utero and the outcome of these pregnancies. STUDY DESIGN: A total of 20 cases with unexplained IUGR and 20 cases with appropriate growth for gestational age has been studied. Amnion, chorion and villi biopsy specimens were obtained from growth-retarded cases and controls at delivery. Cord blood specimens for 48-hour lymphocyte cultures were obtained from all infants with IUGR. RESULTS: Karyotype analysis revealed confined placental mosaicism in two of 20 (10%) cases with IUGR. In one growth retarded case and one appropriate growth for gestational age case, mosaicism was also confirmed in the amnion. Cytogenetic analysis from peripheral blood of newborns showed normal karyotype in all cases. Three pregnancies in the group of fetuses with IUGR (15%) ended with fetal death compared with normal fetal surveillance of all cases from the control group. CONCLUSION: Confined placental mosaicism was detected two times more frequently from placentas of growth- retarded infants compared with those of newborns with appropriate growth. The fetal loss was significantly higher in the group of cases with IUGR compared with the control group.

Prenatal diagnosis of spinal muscular atrophy type I (Werdnig- hoffmann) by DNA deletion analysis of cultivated amniocytes.

AIM: Presentation of a prenatally diagnosed case of Werdnig-Hoffmann disease, the most severe type of spinal muscular atrophy. METHODS: DNA obtained from cultivated amniocytes was analyzed for deletions in the survival motor neuron gene and neuronal apoptosis inhibitory protein gene. RESULTS: The fetus was diagnosed as an affected homozygote for deletions in exon 7 and exon 8 of the survival motor neuron gene. No deletions of exon 5 in the neuronal apoptosis inhibitory protein gene were found. CONCLUSION: Direct DNA deletion analysis of the survival motor neuron gene and neuronal apoptosis inhibitory protein gene in affected families represents a highly reliable and fast method for prenatal diagnosis of Werdnig-Hoffmann disease.

Ultrasonic markers of fetal chromosomal abnormalities.

OBJECTIVE: The aim of this brief investigation was to correlate the most common sonographically detectable markers with certain type of chromosomal disorder diagnosed by available karyotyping procedures. STUDY DESIGN: During the 3 year study period fetal karyotyping was performed in 1055 patients for a variety of clinical indication. Twenty one percent (21%; 222/1055) of procedures were done because of sonographically detectable structural disorders related to phenotype expression of chromosomopathies. Sonographic examinations and karyotyping procedures were performed between the 10th and 36th week. The average maternal age was 27 years, unselected. RESULTS: The fetal karyotype was abnormal in 13.5% of cases (30/222). Within the group of single marker, 11.6% (7/60) of karyotypes were abnormal. Multiple markers of chromosomal abnormalities resulted in 14.2% (23/162) of abnormal karyotypes. The most frequent chromosomal disorder detected in sonographic screening is trisomy 18 (50%; 15/30). The data on the frequency of different types of chromosomal abnormalities are given. CONCLUSIONS: The incidence of chromosomal abnormalities for ultrasonographically detectable malformations is much higher than the incidence reported in screening studies based on maternal age or biochemical screening. Trisomy 21 showed the relative lack of variety in phenotypic expression, and nuchal translucency screening has to be accepted rationally. Associated numerous major and minor malformations were the most prominent factors leading to the diagnosis of chromosomopathies, particularly trisomy 18.

Incidence of chromosomopathies and cystic fibrosis mutations in second trimester fetuses with isolated hyperechoic bowel.

OBJECTIVE: Fetal echoic bowel can be a normal second trimester ultrasonographic finding which usually disappears by 20 weeks on serial sonograms. Recent studies have suggested a possible association of hyperechoic fetal bowel with chromosomopathies and cystic fibrosis. The aim of our study is to determine the incidence of chromosomopathies and cystic fibrosis mutations among the fetuses with isolated hyperechoic bowel. METHODS: Sixteen fetuses with isolated echoic bowel were detected: 13 fetuses < or =20 weeks gestation (group I) and 3 fetuses at 20-26 weeks gestation (group II). Cytogenetic studies were performed in all 16 cases and 11 families had deoxyribonucleic acid-based risk assessment for cystic fibrosis. The echogenity of bowel was that of surrounding bone. RESULTS: Two cases of trisomy 21 and 1 case of trisomy 13 were detected (18.7%). The other ultrasonographic markers begin to appear after 21 weeks gestation in fetuses with trisomy 13. Two of 3 pregnant women with pathological karyotype were younger than 35 years. One of 11 cases (9%) was found to be a heterozygote carrier for deltaF508 mutation. CONCLUSIONS: Isolated hyperechoic bowel in the second trimester was found to be associated with a significantly higher risk of fetal aneuploidy.

Intervillous blood flow in patients with missed abortion.

AIM: Analysis of the color Doppler features of intervillous flow in cases of missed abortion, and their comparison with the findings in normal pregnancy. METHODS: Transvaginal color and pulsed Doppler was used in the analysis of 38 patients with missed abortion and 40 patients with normal pregnancy. The gestational age ranged from 6 to 11 weeks. Repeated analysis of variance was used to compare the groups. RESULTS: Two types of Doppler signals were detected in the intervillous space: pulsatile and continuous. Impedance in the intervillous space of both groups did not change significantly with the gestational age. For the group with missed abortion, the mean resistance index (RI) was 0.37 +/- 0.03 and pulsatility index (PI) 0.75 +/- 0.07. For the control group, the mean RI was 0.36 +/- 0.02 and PI 0. 72 +/- 0.04. The differences in RI and PI for all gestational age groups and between the two patient groups were not statistically significant. With advancing gestation, continuous, venous-like signal became stronger and randomly dispersed throughout the placenta. Peak flow velocity values obtained from both groups did not change significantly with gestational age, nor between the two groups. Conclusion. The new generation of sensitive Doppler units can detect the intervillous flow as a continuous progressive process at 6-11 weeks of gestation. The vascular resistance in the intervillous space does not change in cases of missed abortion.

Histopathologic findings of placenta and arcuate artery Doppler velociometry in preeclampsia and intrauterine growth retardation.

The aim was to analyze the histopathologic changes of placentas and to compare them to the results of arcuate artery color Doppler velocimetry. Fifty four placentas were from pregnancies complicated with pre-eclampsia that ranged from mild forms to convulsions (group 1), 42 from pregnancies complicated with intrauterine growth retardation (IUGR) (group 2) and 40 from uncomplicated pregnancies (group 3). The arcuate artery resistance index (AARI) was increased in 66.66% in group 1 and 59.52% in group 2 (NS). In all uncomplicated pregnancies (group 3) AARI was normal. In group 2, increased AARI was significantly more frequently associated with minimal hypoxic damage (MHD) of placental tissue than in group 1 (p < 0.005), whereas multiple infarcts were more common in group 1 than in group 2 (p < 0.005). At normal AARI multiple infarcts were significantly more frequently found in group 1 than in groups 2 and 3 (p < 0.005), whereas normal placental findings were significantly more common in group 3 than in groups 1 and 2 (p < 0.001). Hypoxic lesions were significantly more often associated with increased AARI (p < 0.01). The positive predictive value of arcuate artery color Doppler velocimetry for hypoxic placental lesions was 93%, and negative predictive value was 10%. Sensitivity and specificity of the method in the prediction of hypoxic placental lesions was 62% and 91%, respectively.

Intervillous circulation in all three trimesters of normal pregnancy assessed by color Doppler.

Our cross-sectional study included 115 healthy pregnant women in gestational age between 7 and 24 weeks. The aim of the study was to compare resistance (RI) and pulsatility (PI) indices, peak systolic velocity (PSV), end-diastolic velocity, and temporal averaged maximum velocity (TAMV) of the spiral arteries and vessels within the intervillous space in all three trimesters of pregnancy. The impedance to blood flow within the intervillous space significantly decreased towards the mid-pregnancy (p < 0.01) and then remained stable. Blood flow velocities within the intervillous space expressed by PSV, EDV and TAMV increased significantly (p < 0.01) towards the mid-pregnancy. After reaching the plateau between 16 and 22 weeks of gestation, these parameters remained almost constant until the 36th gestational week. Near the term low-significant decrease of blood flow velocities was noted: for PSV and TAMV p < 0.07, and for EDV p < 0.05. A significant increase in continuous intervillous blood flow velocity was noted from the 11th week onward (28 +/- 12 vs. 36 +/- 12) until the 36th week of gestation (36 +/- 12 cm/s). The first report of the haemodynamic changes within the intervillous space during pregnancy may have implication in better understanding of the metabolical interchange between maternal and fetal side.

Osteogenic protein-1 is produced by human fetal trophoblasts in vivo and regulates the synthesis of chorionic gonadotropin and progesterone by trophoblasts in vitro.

Osteogenic protein-1 (OP-1/BMP-7), a member of the transforming growth factor (TGF-beta) superfamily of proteins, is shown to be expressed at sites of epithelial-mesenchymal tissue interaction during human fetal development. In the present study, we examined the expression of OP-1 in human placentas (5-11 weeks of gestation and full term) using in situ hybridization and immunohistochemistry methods. The results show that OP-1 was expressed in cytotrophoblasts (Langhans layer) of the chorionic villi in early and full term placentas. Employing highly purified cytotrophoblast cultures which fused to form functional syncytiotrophoblasts, we showed that exogenously added recombinant OP-1, in the presence of low serum concentration, reduced the secretion of chorionic gonadotropin and progesterone by 60% and 36%, respectively, compared with control cultures. The results suggest that osteogenic protein-1 is synthesized locally by cytotrophoblasts and may be involved in the regulation of human pregnancy by controlling reproductive hormonal secretion.

Effect of pulse Doppler ultrasound on placental hormone excretion in vitro.

There is no literature data on the effects of ultrasound on the metabolism of placental tissue. In the present study, the authors have investigated the possible influence of pulsed field Doppler ultrasound on hormonal excretion of human placental trophoblasts in vitro. For the detection of placental hormones, specific immunoassays for chorionic gonadotropin (hCG), human placental lactogen (hPL), estrogen and progesterone were used. Compared to control trophoblast hormone excretion, pulsed field ultrasound exposure induced no changes in the excretion of the hormones in vitro. The authors were unable to demonstrate any alterations caused by pulsed field Doppler ultrasound in this in vitro model. Additional studies are required to deepen our understanding of the effect of pulsed Doppler ultrasound on other biomolecules and their metabolism.

Expression pattern of osteogenic protein-1 (bone morphogenetic protein-7) in human and mouse development.

Osteogenic protein-1 (OP-1; BMP-7) is a member of the bone morphogenetic protein subfamily. Because members of the TGF-beta superfamily have a role in tissue development, the distribution of OP-1 expression in developing human embryos (5-8 gestational weeks) and fetuses (8-14 gestational weeks) and mouse (9.5-17.5 gestational days) fetuses was examined. Northern hybridization with specific OP-1 probes revealed two mRNA species of 4 and 2.2 KB. Highest levels of OP-1 mRNA were found in human fetal kidney and heart between 12-14 weeks of gestation. By in situ hybridization, the OP-1 transcripts were found in various tissues, i.e., the ectodermal epithelium of the mouse fore- and hindlimbs, heart, teeth, intestinal epithelium, perichondrium, hypertrophic chondrocytes, and periosteum/osteoblast layer of developing human bones. In kidneys, transcripts were first detected in the epithelium of the branching uretheric buds, whereas at later stages glomeruli were the major site of OP-1 mRNA accumulation. These data suggest that, although OP-1 has been isolated from bone matrix, it may have additional regulatory roles in the morphogenesis and/or function of the kidney, limb bud, tooth, heart, and intestine.

Glycoprotein and ganglioside changes in human trophoblasts after exposure to pulsed Doppler ultrasound.

Changes in glycoprotein and ganglioside composition in human trophoblasts (eighth week of gestation) after in vitro exposure to pulsed Doppler ultrasound (pulse duration 1.22 microseconds; repetition frequency 11.1 kHz; center frequency 4 MHz; ISPPA = 175.5 W/cm2; ISPTA = 0.59 W/cm2) were investigated. Evacuated trophoblasts were divided in two halves and insonated for 10 min on top of a 6-cm layer of 5% gelatin in 50-mL tubes (Falcon) at 37 degrees C. One half of each trophoblast was sham insonated and served as an internal control. After insonation trophoblasts were maintained at 37 degrees C for 24 h. Glycoproteins were detected using alpha-D-mannose specific lectins from Galanthus nivalis and Narcissus pseudonarcissus. A decrease in the expression of mannose containing glycoprotein mgp47 and an increase in expression of mgp54 were observed. Ganglioside composition was also significantly altered. Concentrations of two gangliosides migrating similarly to GM2, and one similarly to GQ1, decreased by more than 75%. At the same time, concentrations of one ganglioside migrating similarly to GM3, and two other unidentified gangliosides increased two- to fourfold.

Corpus luteum blood flow in normal and abnormal early pregnancy: evaluation with transvaginal color and pulsed Doppler sonography.

One hundred and thirteen (66.5%) women in this study had a normal intrauterine pregnancy with ages ranging 6 to 12 weeks of gestation. Fifty-seven (33.5%) patients were admitted to the hospital owing to clinically suspected abnormal early pregnancy. Dilatation and curettage were done on all women and tissue sample sent to the pathologist for a final diagnostic. Diagnosis of ectopic pregnancy was made on laparoscopy. Both ovaries were examined carefully by color Doppler in sonography in all patients. Color flow was used as a guide for pulsed Doppler exploration. Corpus luteum blood flow was defined as random, usually semilunar in appearance, dispersed vessels with very low impedance to blood flow. The resistive index and pulsatility index were calculated. Overall detection rate of corpus luteum blood flow in normal pregnancies was higher for the left ovary (62.6%) than for the right ovary (37.4%) (P < 0.01). The mean resistive and pulsatility indices from corpus luteum blood flow were not influenced by gestational age in normal pregnancy. The overall mean value for for resistive index was 0.452 +/- 0.04 and for pulsatility index 0.636 +/- 0.09. The overall detection rate of corpus luteum in abnormal pregnancies also was higher for the left ovary (56.7%) than for the right ovary (43.4%) (P < 0.01). The mean resistive indices from corpus luteum blood flow in patients with missed abortion was higher than in women with normal pregnancy (P < 0.01). Both resistive and pulsatility indices were higher in patients with incomplete or threatened abortion in comparison with normal pregnancy (P < 0.01). No statistically significant difference was seen in the case of anembryonic, molar, or ectopic pregnancy.

Antithrombin III in fetuses and newborn children.

Antithrombin III (AT III) was measured in plasma of healthy fetuses between 20-29 weeks of gestation, in plasma of suspected growth retarded or malformed fetuses between 30-39 weeks of gestation, in healthy newborn infants after delivery, and in healthy infants during the first year of life. Measurements were performed with an AT III assay (Orion Diagnostica, Espoo, Finland Turbox) on nephalometer Turbox. The results were expressed as a percentage of the mean adult value (300 mg/l) and statistically analysed with the non-parametric Kruskal-Walles test. AT III levels in fetuses were low but increasing. They continued to increase after birth (F = 34.53 p < 0.001) and reached adult values in the age between the tenth and twelfth month of life.

Localization of osteogenic protein-1 (bone morphogenetic protein-7) during human embryonic development: high affinity binding to basement membranes.

Osteogenic protein-1 (OP-1) is a member of the bone morphogenetic protein subfamily of the transforming growth factor-beta (TGF-beta) superfamily. Since members of the TGF-beta superfamily have a role in tissue development the distribution of OP-1 in developing human embryos (gestational age 5-14 weeks) was examined by immunohistochemical methods. Positive staining for OP-1 was observed in: sclerotome, hypertrophied chondrocytes, osteoblasts, periosteum, epithelial cells of the adrenal "provisional cortex" and the convoluted tubules of developing kidneys. In the developing lungs, pancreas and skin, OP-1 was localized in basement membranes underlying the epithelium. In vitro binding studies of 125I-OP-1 to various extracellular matrix components revealed high affinity of OP-1 for type IV collagen and less for heparin, collagen types I and VI. Present findings suggest that, in addition to bone formation, OP-1 could have other important regulatory roles in human embryogenesis with high binding affinity to a basement membrane component.

[Pregnancy outcome after early amniocentesis]. / Ishod trudnoce nakon rane amniocenteze.

The indications, karyotype, and outcome of 1812 pregnancies following midtrimester amniocentesis (eACZ/as) performed from 1977 to 1989 are presented, especially regarding the risk of pathologic karyotypes and spontaneous abortion. There were 37 pathologic karyotypes (trisomy 21 in 24 pregnancies). The spontaneous abortion rate following amniocentesis was 2.8% although it was almost halved after direct ultrasound guidance had been introduced (1.87% versus 3.66%). In gravidas with previous spontaneous abortion, the preterm delivery rate and spontaneous abortion rates following rACZ were significantly increased. There is a progressive increase of the spontaneous abortion rate after eACZ with the number of previous spontaneous abortions; 5.83% when there was only 1 previous spontaneous abortion, 7.83% when there were 2, and 8.75% when there were 3 or more spontaneous abortions. Previous spontaneous abortions do not increase the risk of chromosomal aberration. In women aged 37 years or more the risk of trisomy 21 is 0.69%, aged 38 years 1.55% and over 39 years 0.44%. The risk is rapidly increasing after 40 years of age (2.25%). The proportion of pathologic karyotypes in these age groups was significantly higher in comparison to younger women. The age limit of 37-38 years as the indication of a possible appearance of mid-trimester ACZ is discussed.

[Correlation between levels of prolactin in maternal serum, fetal serum and amniotic fluid in childbirth at term]. / Korelacija vrednosti nivoa prolaktina u serumu majke, serumu ploda i plodovoj vodi kod porodaja trudnice u terminu.

In 117 parturients with 38-41 gestation weeks, prolactin was analysed by the radioimmunologic method in the mother serum, the umbilical cord serum and vein, and in the amnionic fluid. Three groups of parturients and their newborns were examined: the group (n = 44) with the birth having started spontaneously with the appearance of labour pains 1/10 minutes, the group (n = 38) with a programmed birth induced by the infusion of oxytocin, and the group (n = 35) with the birth comprising elective cesarean section. The concentration of prolactin in the examined sera is characterized by considerable individual oscillations. The highest prolactin level was in the amnionic fluid (395.6 +/- 130.1 mu/L) and the lowest in the mother serum (174.6 +/- 84.1 mu/L) which shows a statistically significant difference (p less than 0.01). Prolactin values in the umbilical artery (244.6 +/- 98.3 mu/L) and vein (230.4 +/- 91.7 mu/L) are significantly (p less than 0.001) higher than the value in the mother sera and significantly lower (p less than 0.001) than the prolactin concentration in the amnionic fluid. The difference of the prolactin values in the sera of the umbilical cord blood vessels has no statistical significance (p greater than 0.05). Nor is there any statistically significant difference between prolactin concentrations in spontaneous and induced deliveries versus those in deliveries terminated with elective cesarean section (p greater than 0.05). A correlation analysis of the functional connection of prolactin in the sera of the mother, fetus, and amnionic fluid gives the correlation coefficient values of high statistical significance (0.482 less than r less than 0.906; p less than 0.001).

[The effect of segmental epidural analgesia on the duration of labor in spastic dystocia]. / Utjecaj segmentalne epiduralne analgezije na trajanje porodaja sa spasticnom distocijom.

In 77 parturients, owing to spastic distotia, the labour was conducted in continuous epidural analgesia, while in the control group of parturients with the same diagnosis (N = 32), the intramuscular use of pethidine and diazepam was repeatedly applied. The epidural catheter was placed at the 2-3 cm dilated cervix. Carticain with or without fentanyl was used as a local anaesthetic. The average duration of labour from the beginning to the 2-3 cm cervical opening was 9.3 hours in the group of epidural analgesia applied parturients and 4.3 hours in the control group. The continuation of labor in the examined group lasted 3.7 hours and in the control group 12 hours. The difference is significant (p less than 0.01). A spontaneous vaginal delivery in the examined group was recorded in 77.9% women. There were 14.3% cesarean sections in the examined and 46.8% in the control group. A protracted labour was a significantly more frequent indication (p less than 0.01) for such a termination of labour in the control group.