Abstract. Brominated flame retardants (BFRs) are used to prevent combustion in consumer products. Examples of BFRs are polybrominated diphenyl ethers (PBDEs), tetrabromobisphenol A (TBBPA), and tribromophenol (TBP). These compounds are reported to have adverse effects on human health and endocrine disrupting effects. The purpose of this study was to identify the Japanese perinatal exposure to PBDEs, hydroxylated PBDE metabolites (OH-PBDEs), TBBPA, and TBP compared with polychlorinated biphenyls (PCBs) and hydroxylated PCB metabolites (OH-PCBs). We investigated the concentrations of these compounds in maternal blood, maternal milk, cord blood, and umbilical cords from 16 Japanese mother-infant pairs by HRGC/HRMS. PBDEs were detected in all samples of maternal blood (mean ± SD; median = 25 ± 23 pg/g; 18 pg/g wet weight), maternal milk (140 ± 220 pg/g; 59 pg/g wet weight), cord blood (4.8 ± 6.5 pg/g; 1.6 pg/g wet weight), and umbilical cords (3.1 ± 3.1 pg/g; 2.1 pg/g wet weight). The mothers were divided into two groups, a high-concentration group and a low-concentration group. The percentage of BDE-47 showed the greatest difference between the two groups. 6-OH-BDE-47, TBBPA, and TBP were detected in all umbilical cord samples (mean ± SD; median = 8.4 ± 8.1 pg/g; 8.0 pg/g, 16 ± 5.5 pg/g; 15 pg/g, and 33 ± 8.2 pg/g; 32 pg/g wet weight respectively), but not in all maternal blood or cord blood samples. These results indicate that OH-PBDEs, TBBPA, and TBP, in addition to PBDEs, PCBs, and OH-PCBs, pass through the blood-placenta barrier and are retained in the umbilical cord.
Three types of tissue samples—umbilical cord (UC), umbilical cord serum (CS), and maternal serum (MS)—have often been used to assess fetal exposure to chemicals. In order to know the relationship of contamination between mothers and fetuses, we measured persistent chemicals in comparable sets of the three tissue samples. Also, we analyzed the association between the chemicals in maternal and fetal tissues to know which tissue is the best sample for fetal exposure assessment. On a wet basis, the chemical concentrations were of the order MS > CS > UC, except for some chemicals such as cis-chlordane and endosulfan. On a lipid basis, the concentrations in UC were nearly equal or often higher than in MS, but the concentrations in CS were usually lower than in others. Hexachlorocyclohexanes and penta-, hexa-, and heptachlorinated biphenyls showed an association between the concentrations in UC versus MS, and UC versus CS. These chemicals also showed high correlation coefficients between the chemical concentrations in UC of first babies and maternal age. These chemicals were closely related to each other when grouped on the basis of their concentrations using cluster analysis. In conclusion, we insist that UC is the best sample to assess fetal contamination status of persistent chemicals. There is a possibility that the assessment based on the contamination levels in CS result in an underestimation.
The determination of bisphenol A (BPA) and/or bisphenol A diglycidyl ether (BADGE) in foods sold in Japanese markets and in water leached from six epoxy resin cans with similar diameters was carried out using high-performance liquid chromatography (HPLC) with electrochemical detection (LC/ECD), LC-mass spectrometric detection (LC/MS) and LC-tandem mass spectrometric detection (LC/MS/MS). BPA concentrations were 0-842 ng g(-1) for 48 canned foods, 0-14 ng g(-1) for 23 foods in plastic containers, and 0-1 ng g(-1) for 16 foods in paper containers. No BADGE was detected in three canned foods. There was no difference in leaching concentrations of BPA into glycine buffers at pHs 8 and 11, and water. The amounts of BPA leached into water from six epoxy resin cans held at 121 degrees C for 20 min were almost the same as the cans' contents and were much higher than the amounts leached from cans held at or below 80 degrees C for 60 min. The amount leached depended on the type of can, but not on the amount of BADGE leached from the cans. Considerably more BPA than BADGE leached to water from six cans. Two cans whose contents had high concentrations of BPA showed no BADGE leaching even at 121 degrees C, suggesting the different kinds of epoxy resin can linings from others. The results imply that the main source of human exposure to BPA is food from cans with linings that contain high percentages of BPA as an additive or an unforeseen contaminant.
Abstract. Perinatal exposure to diethylstilbestrol (DES) can have numerous adverse effects on the reproductive organs later in life, such as vaginal clear-cell adenocarcinoma. Epigenetic processes including DNA methylation may be involved in the mechanisms. We subcutaneously injected DES to neonatal C57BL/6 mice. At days 5, 14, and 30, expressions of DNA methyltransferases (Dnmts) Dnmt1, Dnmt3a, and Dnmt3b, and transcription factors Sp1 and Sp3 were examined. We also performed restriction landmark genomic scanning (RLGS) to detect aberrant DNA methylation. Real-time RT-PCR revealed that expressions of Dnmt1, Dnmt3b, and Sp3 were decreased at day 5 in DES-treated mice, and that those of Dnmt1, Dnmt3a, and Sp1 were also decreased at day 14. RLGS analysis revealed that 5 genomic loci were demethylated, and 5 other loci were methylated by DES treatment. Two loci were cloned, and differential DNA methylation was quantified. Our results indicated that DES altered the expression levels of Dnmts and DNA methylation. [3][4][5], and DES is now widely known as an estrogenic endocrine disruptor. In our previous studies, we showed that mice treated neonatally with DES or the phytoestrogen, genistein, developed altered expression levels of various genes in male reproductive organs, even during adulthood [6][7][8][9][10]. Since the expression changes lasted for long periods of time, and DES was reported to be non-genotoxic [11], epigenetic mechanisms might be involved in the gene expression changes. DNA methylation has an important role in epigenetics, and the DNA methy-
Several types of methods, mainly liquid chromatography (LC), have been used for the analysis and assessment of bisphenol A (BPA) in human biological samples. Enzyme-linked immunosorbent assay (ELISA) is also being used for BPA measurement. In this study, we verified whether ELISA is suitable for measuring human samples, namely, serum and urine, by comparing the ELISA results with those obtained by liquid chromatography with multichannel colometric electrochemical detection (LC/ECD) and liquid chromatography coupled to a mass spectrometer (LC/MS/MS). Results of the measurement with LC/ECD showed urinary BPA concentrations to be 1.92 [1.45] +/- 1.99 (mean [median] +/- standard deviation) ng BPA/mL without the correction of urine volume and 1.20 [0.90] +/- 1.10 mug BPA/g creatinine; however, in serum, free and total BPA were not detected. In both samples, a good correlation of the values with the methods was not found. ELISA is one of the powerful measurement methods, since it is convenient and useful for screening bulk quantities. At this point, however, ELISA is not suitable for BPA measurement in human samples because of low levels of BPA in human samples, matrix effect, and specificity of anti-BPA antibody.
Abstract.We investigated the long-term estrogenic influence of genistein on the male reproductive system in mice. Newborn ICR male mice were treated with genistein (10, 100, or 1000 , tg/mouse) for neonatal 5 days. As positive control, administration of diethylstilbestrol (0.5-50 pg/mouse) was carried out. In mice exposed to genistein,we examined weight of testes, sperm counts, sperm motility, and mRNA expression levels of estrogen receptor a (ERa) and androgen receptor (AR) at 4, 8 or 12 weeks after birth. Moreover, at 12 weeks of age, we evaluated protein level of ERa. In our conventional reproductive-toxicological study (weight of testes, sperm counts and sperm motility), neonatal transient exposure to genistein did not show adverse effects on the male reproductive system in 4, 8 or 12 week old mice. However, in mice treated with genistein mRNA expression levels of ERa and AR were reduced at 8 weeks. This reduction was recovered at 12 weeks in mice treated with a lower dose (10 ICg) of genistein but not in those with higher doses (100 pg and 1000 rig). In addition, ERa protein levels tended to decrease in 12 weeks of adulthood.Our results exhibited that the disruption of gene expression continued for long term such as 3 months after administration of genistein, even if no effect was found at conventional reproductive-toxicological level. We have shown that neonatal administration of weak estrogenic compound (genistein) affects male reproductive organs at molecular levels in adulthood.
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