Effects of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) on preimplantation mouse embryos

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent environmental contaminant that can exert developmental toxicity. To investigate the stage-specific effects of TCDD on preimplantation embryos, we exposed mouse embryos to TCDD at different stages (1-, 2-, and 8-cell) and collected them at different stages of development (the 1- or 2-, 8-cell, and blastocyst stage, respectively). Semiquantitative RT-PCR revealed increased constitutive gene expression of the arylhydrocarbon receptor (AhR) and AhR nuclear translocator (Arnt) at the 1-cell stage, decreased expression at the 2- to 8-cell stage, and increased expression again at the blastocyst stage, and addition of TCDD to media did not affect their mRNA levels. Interestingly, no cytochrome P4501A1 (CYP1A1) mRNA was detected in embryos at the 1-, 2-, and 8-cell stages after exposure to 10 nM TCDD for 12 or 24 h, whereas CYP1A1 mRNA was significantly increased at the blastocyst stage in response to TCDD, and its induction was found to be concentration-dependent on TCDD exposure from 0.01 to 10 nM for 24 h. In addition, no significant differences in development rate of preimplantation embryos, cell number of blastocyst embryos, or apoptotic indices, such as TUNEL-positive cell number or Bax/Bcl-2 expression ratios were observed at the blastocyst stage between TCDD-exposed groups and non-exposed group. These results suggest that the sensitivity to TCDD differs with the embryonic stage, which may reflect an ability of embryos to adapt to environmental stressors, such as dioxins.     

Increased glycogen content and glucose transporter 3 mRNA level in the placenta of Holtzman rats after exposure to 2,3,7,8-tetrachlorodibenzo-p-dioxin

Exposure to a low dose of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) produces a variety of toxic manifestations, including fetal death. In order to evaluate the effects of low-dose TCDD on placental function in this study, pregnant Holtzman rats were given a single oral dose of 800 or 1600 ng TCDD/kg body wt or an equivalent volume of vehicle (control) on gestation day (GD) 15 and the results were observed on GD16 and GD20. The number of fetal deaths increased in the animals exposed to TCDD. Although fetal and placental weight did not differ significantly between the control group and the TCDD groups, histological differences from the control rats were clearly observed in the junctional zone (JZ) of the placentas of the TCDD-exposed rats. In the control placenta, glycogen cells occupied the majority of the JZ on GD16, but then decreased in number and almost disappeared by GD20, whereas on GD20 the placenta of the TCDD-exposed rats exhibited a larger area occupied by the glycogen cells and cysts filled with eosinophilic material surrounded by glycogen cells in the JZ than that of the control group. Glycogen assay revealed that the glycogen content of the placentas from the TCDD-exposed rats was higher than in the control rats. Semiquantitative RT-PCR analysis was performed to assess the expression of glucose transporter 1 (GLUT1) and GLUT3, the two major placental glucose transporter isoforms. On GD20 the level of expression of GLUT1 mRNA in the placentas was not different between the control and TCDD groups, whereas the level of expression of GLUT3 mRNA approximately doubled in both the 800 and 1600 ng/kg TCDD groups. GLUT3 mRNA expression was restricted to the labyrinth zone of placenta, where zone-specific expression of mRNA arylhydrocarbon receptor and induction of cytochrome P450 1A1 mRNA by TCDD were observed, and none was detected in the JZ. These results, including the increase of glycogen content and GLUT3 mRNA level in TCDD-exposed placentas, provide the first evidence of alteration of glucose kinetics in the placenta by TCDD.     

Depth scaling of solid phantom for intensity modulated radiotherapy beams

To reduce the uncertainty of absorbed dose for high energy photon beams, water has been chosen as a reference material by the dosimetry protocols. However, solid phantoms are used as media for absolute dose verification of intensity modulated radiotherapy (IMRT). For the absorbed dose measurement, the fluence scaling factor is used for converting an ionization chamber reading in a solid phantom to absorbed dose to water. Furthermore the depth scaling factor is indispensable in determining the fluence scaling factor. For IMRT beams, a photon energy spectrum is varied by transmitting through a multileaf collimator and attenuating in media. However, the effects of spectral variations on depth scaling have not been clarified yet. In this study, variations of photon energy spectra were determined using the EGS Monte Carlo simulation. The depth scaling factors for commercially available solid phantoms were determined from effective mass attenuation coefficients using photon energy spectra. The results clarified the effect of spectral variation on the depth scaling and produced an accurate scaling method for IMRT beams.     

Determination of dioxins in human hair: Estimation of external and internal exposure to dioxins

Objectives To clarify the origin of dioxin and related compounds (dioxins) in human hair, we determined the amounts of adsorbed dioxins in human hair, and the distribution of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) in rats. Methods Human hair specimens, packed in a glass column, were exposed to ambient air that was introduced into the column with an air pump for 24 h. Rats were administered TCDD by gavage at doses of 0.2, 0.8, and 1.6 μg/kg body weight. Four weeks after TCDD administration, hair from the back, serum, and adipose tissue were removed under diethyl ether anesthesia. The amounts of dioxins in these samples were analyzed by high resolution gas chromatography with mass spectroscopy. Results Exposure of the hair specimens to ambient air for one day increased the total toxic equivalent (TEQ) value by 51%. In TCDD-treated rats, the amount of TCDD in hair increased in a dose-dependent manner, and showed a significant positive correlation with that in adipose tissue. Conclusions Human hair was found to retain dioxins by both internal and external exposure, and the contribution of external exposure was estimated to be about 40% of the TEQ.     

Expression of basic fibroblast growth factor receptor messenger RNA in the periinfarcted brain tissue

The present study examined whether expression of basic fibroblast growth factor receptor (bFGFR) messenger ribonucleic acid (mRNA) was upregulated by focal ischemia. We have studied the in situ hybridization autoradiography for bFGFR mRNA in the rat model of middle cerebral artery (MCA) occlusion. Male Wistar rats were used for occlusion of the left MCA, and were sacrificed 1, 3, 7 and 14 days after MCA occlusion. In situ hybridization was performed on the brain sections of these animals and sham controls by using 35S-labeled antisense and sense (control) RNA probes for rat bFGFR. Expression of bFGFR mRNA was observed in the periinfarcted area of the rats within 1-14 days after MCA occlusion. Expression was evident in the whole hemisphere of the infarcted side, especially at 1 and 3 days after ischemia, but no expression was detected in the contralateral side. On microautoradiograms, the signals of bFGFR mRNA were detected in both neurons and non-neural cells located in the periinfarcted area. Upregulation of bFGFR mRNA detected in the periinfarcted brain tissue suggests that receptor-mediated action of bFGF may be related to preservation of neurons injured by ischemia.     

Study of Electrical Power Facilities and Measures for Planned Outages in Japanese Hemodialysis Clinics After the Great East Japan Earthquake

The Great East Japan Earthquake on 11 March 2011 caused major damage in northeastern Japan. The Kanto region experienced a massive electrical power shortage in the summer of 2011. A questionnaire was submitted to 354 hemodialysis clinics in Kanagawa prefecture and the Tokyo metropolitan area, excluding isolated islands, and 176 responses were analyzed (49.7%). The questions included evaluation of the availability of a private electricity generator, countermeasures in case of a planned outage, awareness of saving electricity, and improvement of safety of medical devices or electrical facilities after the earthquake. Only 12% of the clinics had private electricity generators and many clinics had no plans to introduce this facility. However, 96% of the clinics had established countermeasures to deal with a planned outage. Many clinics planned to provide dialysis on a different day or at a different time. All clinics had tried hard to establish procedures to save electricity in the summer of 2011, and 84% of the clinics had reconsidered and improved the safety of medical devices or electricity facilities after the earthquake. These results show that the awareness of crisis management was greatly improved in the wake of the earthquake.