Effects of diagnostic ultrasound on HSP70 expression in chorionic villi in rats during early pregnancy and the role of HSP70 in apoptosis in chorionic villi.

The aim of this study was to investigate the effects of diagnostic ultrasound on chorionic villi during early pregnancy in rats through the examination of heat shock protein 70 (HSP70) expression and apoptosis in chorionic villi. Using TUNEL, RT-PCR, western blot analysis and immunohistochemistry, the extent of apoptosis and the gene/protein expression of HSP70, Bcl-2 and Bax was determined in the samples of the chorionic villus from the rats during early pregnancy. Compared with the unexposed group, there were clear signs that apoptosis had occurred in the villi exposed to ultrasound for 30 min. The expression of HSP70 and Bax in the villi gradually increased with the extended duration of exposure to ultrasound, whereas the expression of Bcl-2 gradually decreased. Bcl-2 expression did not differ significantly between the group exposed for 10 min and the group exposed for 20 min. Taken together, our data demonstrate that HSP70 may protect chorionic villi exposed to ultrasound by inhibiting apoptosis; however, its protective effects are limited over the extended duration of exposure to ultrasound. Thus, the prudent use of ultrasound during the early stages of pregnancy should be advocated.

Proteomic analysis on the alteration of protein expression in the early-stage placental villous tissue of electromagnetic fields associated with cell phone exposure.

BACKGROUND: To explore the possible adverse effects and search for cell phone electromagnetic field (EMF)-responsive proteins in human early reproduction, a proteomics approach was employed to investigate the changes in protein expression profile induced by cell phone EMF in human chorionic tissues of early pregnancy in vivo. METHODS: Volunteer women about 50 days pregnant were exposed to EMF at the average absorption rate of 1.6 to 8.8 W/kg for 1 hour with the irradiation device placed 10 cm away from the umbilicus at the midline of the abdomen. The changes in protein profile were examined using 2-dimensional electrophoresis (2-DE). RESULTS: Up to 15 spots have yielded significant change at least 2- to 2.5-folds up or down compared to sham-exposed group. Twelve proteins were identified- procollagen-proline, eukaryotic translation elongation factor 1 delta, chain D crystal structure of human vitamin D-binding protein, thioredoxin-like 3, capping protein, isocitrate dehydrogenase 3 alpha, calumenin, Catechol-O-methyltransferase protein, proteinase inhibitor 6 (PI-6; SerpinB6) protein, 3,2-trans-enoyl-CoA isomerase protein, chain B human erythrocyte 2,3-bisphosphoglycerate mutase, and nucleoprotein. CONCLUSION: Cell phone EMF might alter the protein profile of chorionic tissue of early pregnancy, during the most sensitive stage of the embryos. The exposure to EMF may cause adverse effects on cell proliferation and development of nervous system in early embryos. Furthermore, 2-DE coupled with mass spectrometry is a promising approach to elucidate the effects and search for new biomarkers for environmental toxic effects.

Effects of 50-Hz magnetic field exposure on hormone secretion and apoptosis-related gene expression in human first trimester villous trophoblasts in vitro.

Evidence from epidemiological and animal studies showed that exposure to extremely low frequency magnetic fields (ELF-MF) could produce deleterious effects on reproduction. In order to investigate the possible mechanism of MF exposure on reproductive effects, first trimester human chorionic villi at 8-10 weeks' gestation were obtained, and trophoblasts were isolated, cultured, and exposed to a 50-Hz MF for different durations. The human chorionic gonadotropin (hCG) and progesterone in the culture medium was measured by electrochemiluminescence immunoassay. The mRNA levels of apoptosis-related genes bcl-2, bax, caspase-3, p53, and fas in trophoblasts were analyzed using real-time RT-PCR. The results showed that exposure of trophoblasts to MF at 0.2 mT for 72 h did not affect secretion of hCG and progesterone from these cells. There was also no significant change in secretion of these hormones when trophoblasts were exposed to a 0.4 mT MF for 48 h. However, MF significantly inhibited hCG and progesterone secretion of trophoblasts after exposure for 72 h at 0.4 mT. Results of apoptosis-related gene expression analysis showed that, within 72 h of exposure at 0.4 mT, there was no significant difference between MF exposure and control on the expression pattern of each gene. Based on results of the present experiment, it is suggested that exposure to MF for a longer duration (72 h) could inhibit secretion of hCG and progesterone by human first trimester villous trophoblasts, however, the effect might not be related to trophoblast apoptosis.

[Noise magnetic fields mitigates the inhibition of secretion function of primary human villous trophoblasts induced by 50 Hz magnetic fields].

OBJECTIVE: To explore the effects of 50 Hz sinusoidal magnetic fields (MF) on secretion function of primary human villous trophoblasts in vitro, and the interference effect of "noise" MF. METHODS: The trophoblasts were isolated from human villus by trypsin digestion and incubated in DMEM medium.Then the trophoblasts were exposed to 0.4 mT 50 Hz MF and/or "noise" MF respectively for different durations. Each exposure group was matched with one control group which was from the same villus and cultured with the same condition except the MF exposure. The concentrations of human chorionic gonadotropin (HCG) and progesterone in the culture medium were measured by immunofluorescence. Statistical significance of differences between means was determined by one way-ANOVA with P<0.05 considered significant. RESULT: 50 Hz MF inhibited the HCG and progesterone secretion significantly when exposure for 72 h (compared with control group, P<0.05). There was no significant change of HCG and progesterone secretion when trophoblasts were exposed to 0.4 mT "noise" MF within 72 h (compared with control group, P>0.05). However, by superimposing the "noise" MF, the inhibition of HCG and progesterone secretion of trophoblasts induced by 50 Hz MF was eliminated. CONCLUSION: The exposure to 50 Hz MF for long period could inhibit trophoblasts secreting HCG and progesterone, and the "noise" MF with the same intensity could eliminate the effects induced by 50 Hz MF.

[Oxygen transfer and consumption in human placenta exposed to variable magnetic fields in vitro]. / Transfer i zuzycie tlenu w lozysku ludzkim in vitro w warunkach oddzialywania zmiennego pola magnetycznego.

The initial investigations concerning the influence of variable magnetic fields (MF) on transfer and oxygen consumption in isolated human cotyledon in vitro were performed. Ten dual closed perfusion of the human cotyledon were conducted in each group. The control group was not exposed to magnetic fields. In studied groups cotyledons were exposed to magnetic field: in the group B1 (B = 2 mT, f = 50 Hz), and in the group B2 (B = 5 mT, f = 50 Hz) for 180 min. Obtained results may suggest that variable magnetic field (B = 5 mT, f = 50 Hz) cause decrease of oxygen consumption in human placenta in 120 and 150 min of experiment.

The effect of oscillating low intensity magnetic field on the Na+, K+, Ca++, and Mg++ concentrations in the maternal and fetal circulation of the dually perfused human placental cotyledon.

Dual-sided perfusions of the human placental cotyledon in vitro were used to study effects of low intensity magnetic fields (MFs) of 2 mT, 50 Hz (E1, 10 perfusions) and 5 mT, 50 Hz (E2, 10 perfusions). In the control group C (10 experiments) no field was used. Perfusions lasted 180 min each. Increased release of calcium ions from the placental cotyledon was found in the fetal circulation during perfusion when the 2 mT, 50 Hz MF was used. No changes in the release of sodium and magnesium ions were observed compared to the control group. The 5 mT, 50 Hz oscillating MF intensified the release of sodium ions from the perfused cotyledon both to the fetal and maternal circulation up to the 150th min of the experiment. Increased release of magnesium ions was observed only to the fetal circulation between 120 and 180 min and of calcium ions to the fetal circulation between 60 and 180 min. No significant differences in K concentrations were found between the control and MF exposed cotyledons under conditions of these experiments.

[Evaluation of the morphology of the human placental cotyledon following dual in vitro perfusion in variable magnetic field]. / Ocena morfologii zrazika lozyska ludzkiego poddanego dwustronnej perfuzji in vitro w warunkach zmiennego pola magnetycznego.

UNLABELLED: Objectives and the aim of the study was electron-microscopy morphological estimation of the human placental cotyledon after 180 minutes of dual closed perfusion in vitro. MATERIALS AND METHODS: In the experimental group the cotyledons were exposed to variable magnetic field of 2 mT magnetic induction and 50 Hz frequency. The control group K (10 perfusions) was not subjected to magnetic field while the experimental group B (10 perfusions) was influenced by magnetic field. RESULTS: It was found that homogeneous variable magnetic field disturbs the ultrastructure of the nuclei and cytoplasma and it increases the density of the vascular-epithelial membrane of villi cells of human placenta in vitro. CONCLUSION: Variable sinusoildal, magnetic field of 2 mT magnetic induction and 50 Hz frequency disturbs the ultrastructure of the nuclei and cytoplasma and it increases the density of the vascular-epithelial membrane of villi cells of human placenta in vitro after 180 minutes of dual closed perfusion in vitro.

Prenatal diagnosis of ataxia-telangiectasia and Nijmegen Breakage syndrome by the assay of radioresistant DNA synthesis.

Prenatal diagnosis was performed in 16 pregnancies at risk of ataxia-telangiectasia (A-T) or Nijmegen Breakage Syndrome (NBS). Radioresistant DNA synthesis (RDS) was investigated in cultured chorionic villus (CV) cells and/or amniotic fluid (AF) cells. In four pregnancies, an affected foetus was diagnosed with increased RDS in cultured CV cells. In three of the four cases confirmation of the diagnosis was obtained by analysis of AF cells and/or skin fibroblasts from the foetus cultured after termination of the pregnancy; in the fourth case a fibroblast culture from the aborted foetus failed. In one case, only AF cells could be analysed in a late stage of pregnancy; pregnancy was terminated due to intermediate/equivocal results but the foetal fibroblasts showed normal RDS. Normal RDS was demonstrated in the other 11 pregnancies at 25% risk either by analysis of CV cells (nine cases) or of AF cells (two cases). In some cases the (normal) results on the CV cells were corroborated by subsequent analysis of AF cells. The results suggest that RDS analysis of CV cells allows reliable prenatal diagnosis of A-T/NBS. However, amniocentesis may be necessary to confirm normal results on CV cells if the foetus is female (because of the risk of maternal cell contamination) or in the rare case of equivocal results.

Frequency of chromosomal aberrations after exposure to gamma-radiation of human chorionic villi.

We investigated the chromosomal damage induced by in vitro exposure to gamma-rays of uncultured first trimester chorionic villi. Frequency and types of chromosomal aberrations at increasing doses of radiation have been evaluated on cytotrophoblast spontaneous metaphases obtained after a short term incubation. Our results indicate a direct correlation between radiation dose and aberration frequency.

Spontaneous and induced chromosome breakage in chorionic villus samples: a cytogenetic approach to first trimester prenatal diagnosis of ataxia telangiectasia syndrome.

Patients with ataxia telangiectasia (AT) syndrome exhibit a high level of spontaneous chromosome aberrations, with hypersensitivity to gamma radiation and radiomimetic chemicals at the chromosomal and cellular level. Previously pregnancies at risk for AT have been screened solely by analysis of amniotic fluid samples. In this report we describe a cytogenetic approach to the prenatal diagnosis of AT using chorionic villus sampling (CVS). Levels of spontaneous and induced (gamma radiation and bleomycin) chromosome breakage were established in direct, semidirect, and culture preparations of CVS samples from normal pregnancies. The methods developed were then successfully applied to the screening of a pregnancy at risk for AT. Semidirect preparations showed normal levels of chromosome breakage, and this result was further confirmed in chorion, amniotic fluid, and lymphocyte cultures. In chorion villus samples, gamma radiation is probably the easiest and most reliable way of discriminating between unaffected fetuses and those with AT.