Evaluation of Environmental Contamination and Estimated Radiation Doses for the Return to Residents’ Homes in Kawauchi Village,Fukushima Prefecture

To evaluate the environmental contamination and radiation exposure dose rates due to artificial radionuclides in Kawauchi Village, Fukushima Prefecture, the restricted area within a 30-km radius from the Fukushima Dai-ichi Nuclear Power Plant (FNPP), the concentrations of artificial radionuclides in soil samples, tree needles, and mushrooms were analyzed by gamma spectrometry. Nine months have passed since samples were collected on December 19 and 20, 2011, 9 months after the FNPP accident, and the prevalent dose-forming artificial radionuclides from all samples were ¹³⁴Cs and ¹³⁷Cs. The estimated external effective doses from soil samples were 0.42–7.2 µSv/h (3.7–63.0 mSv/y) within the 20-km radius from FNPP and 0.0011–0.38 µSv/h (0.010–3.3 mSv/y) within the 20–30 km radius from FNPP. The present study revealed that current levels are sufficiently decreasing in Kawauchi Village, especially in areas within the 20- to 30-km radius from FNPP. Thus, residents may return their homes with long-term follow-up of the environmental monitoring and countermeasures such as decontamination and restrictions of the intake of foods for reducing unnecessary exposure. The case of Kawauchi Village will be the first model for the return to residents’ homes after the FNPP accident.     

TRAP1 Controls Mitochondrial Fusion/Fission Balance through Drp1 and Mff Expression

Mitochondria are dynamic organelles that change in response to extracellular stimuli. These changes are essential for normal mitochondrial/cellular function and are controlled by a tight balance between two antagonistic pathways that promote fusion and fission. Although some molecules have been identified to mediate the mitochondrial fusion and fission process, the underlying mechanisms remain unclear. Tumor necrosis factor receptor-associated protein 1 (TRAP1) is a mitochondrial molecule that regulates a variety of mitochondrial functions. Here, we examined the role of TRAP1 in the regulation of morphology. Stable TRAP1 knockdown cells showed abnormal mitochondrial morphology, and we observed significant decreases in dynamin-related protein 1 (Drp1) and mitochondrial fission factor (Mff), mitochondrial fission proteins. Similar results were obtained by transient knockdown of TRAP1 in two different cell lines, SH-SY5Y neuroblastoma cells and KNS-42 glioma cells. However, TRAP1 knockdown did not affect expression levels of fusion proteins. The reduction in Drp1 and Mff protein levels was rescued following treatment with the proteasome inhibitor MG132. These results suggest that TRAP1 regulates the expression of fission proteins and controls mitochondrial fusion/fission, which affects mitochondrial/cellular function.     

Behavioral rhythms of the Japanese newts, Cynops pyrrhogaster, under a semi-natural condition

 Locomotor activity rhythms of the Japanese newt, Cynops pyrrhogaster, were recorded under a semi-natural condition using phototransistor systems. The daily activity rhythm showed a seasonal change: the locomotor activity was mainly diurnal (active during the daytime) from spring to early summer; mainly nocturnal (active during the night-time) from summer to autumn; and showed either a diurnal or nocturnal pattern, depending on the ambient temperature, in winter. To analyze the daily activity in detail, we observed the behavior of a group of newts (three males, three females) throughout 24 h. Four types of behavior (respiration, feeding, mating, and resting on the land) were observed. Each behavior had daily rhythms and showed a seasonal change. The behavior on land showed mainly a nocturnal or bimodal pattern (activity rhythms with two peaks) throughout the year and was more frequently observed in summer. Mating behavior also showed a seasonal change: high activity in spring, with peaks in the early morning and evening, but no activity in summer. Except in winter, feeding and respiratory behavior showed no seasonal changes in either activity period or frequency. Coupling between behavior and the clock seems to be weak in the Japanese newt because of indistinct daily rhythms and frequent phase changes of locomotor activity in water. Physical factors such as humidity and temperature seem to affect strongly the daily activity of the newts. Received: 21 April 1997 / Accepted: 1 September 1997     

PICOPHYTOPLANKTON BIOMASS IN RELATION TO LAKE TROPHIC STATE AND THE TN:TP RATIO OF LAKE WATER IN JAPAN1

Picophytoplankton biomass and its contribution to total phytoplankton biomass were investigated in relation to the nutrient concentration and total N: total P ratio of the epilimnetic waters of 42 Japanese lakes during the warm season in 1991 (April–October). Picophytoplankton biomass (as chlorophyll a) in meso-, eu-, and hypertrophic lakes was significantly higher than those observed in oligotrophic lakes. However, picophytoplankton biomass increased significantly with increased total P concentrations in all systems excluding hypertrophic lakes. Picophytoplankton contribution to total chlorophyll a content was significantly higher in oligo- and mesotrophic lakes than in eu- and hypertrophic lakes and was inversely correlated with total P concentrations in lake water. Picophytoplankton contribution to the total phytoplankton biomass was positively (r = 0.54, n = 42, P = 0.0003) correlated with the total N: total P ratio of lake waters. Each lake trophic type, with the exception of hypertrophic lakes, showed this trend, although the correlation was not significant. We suggest that picophytoplankton contribution is influenced by the total N: total P ratio rather than lake trophic state; however, picophytoplankton were of little importance in hypertrophic lakes.     

Proteolytic processing of phospholipase Czeta and [Ca2+]i oscillations during mammalian fertilization

Phospholipase Cζ (PLCζ) is a sperm-specific PLC capable of causing repetitive intracellular Ca²⁺ ([Ca²⁺]_i) release ([Ca²⁺]_i oscillations) in mammalian eggs. Accumulating evidence suggests that PLCζ is the sperm factor responsible for inducing egg activation. Nevertheless, some sperm fractions devoid of 72-kDa PLCζ showed [Ca²⁺]_i oscillation-inducing and PLCζ-like PLC activity (Kurokawa et al., (2005) Dev. Biol. 285, 376-392). Here, we report that PLCζ remains functional after proteolytic cleavage at the X-Y linker region. We found that N-terminal (33 and 37 kDa) and C-terminal fragments (27 kDa), presumably the result of PLCζ cleavage at the X-Y linker region, were present in fresh sperm as well as in sperm extracts and remained associated as functional complexes. Protease V8 cleaved 72-kDa PLCζ into 33/37 and 27 kDa fragments, while PLC activity and [Ca²⁺]_i oscillation-inducing activity persisted until degradation of the fragments. Immunodepletion or affinity depletion of these fragments abolished PLC activity and [Ca²⁺]_i oscillation-inducing activity from sperm extracts. Lastly, co-expression of cRNAs encoding residues 1-361 and 362-647 of mouse PLCζ, mimicking cleavage at the X-Y linker region, induced [Ca²⁺]_i oscillations and embryo development in mouse eggs. Our results support the hypothesis that PLCζ is the sole mammalian sperm factor and that its linker region may have important regulatory functions during mammalian fertilization.     

Specific apoptosis induction in human papillomavirus-positive cervical carcinoma cells by photodynamic antisense regulation

Human papillomavirus type 18 (HPV18) is frequently detected in cervical cancer cells. The viral proteins E6 and E7 are expressed consistently and have oncogenic activities. The E7 protein binds to a tumor suppressor, the retinoblastoma gene product (pRB), however, leading to the stabilization of tumor suppressor, p53 protein. On the other hand, another viral product, E6, forms complexes with p53 and abrogates its function, resulting in tumor progression. These facts imply that the E6 oncogene is one of the ideal targets for directed gene therapy in HPV-positive cervical cancer. In this study, we tried photodynamic antisense regulation of the antiapoptotic E6 expression using a photocross-linking reagent, 4,5',8-trimethylpsoralen, conjugated oligo(nucleoside phosphorothioate) (Ps-S-Oligo). This photodynamic antisense strategy effectively elicited the apoptotic death of HPV18-positive cervical cancer cells through the selective repression of E6 mRNA and consequent stabilization of p53 protein. E7-mediated signals potentially activated the p53 function and mobilized the p53 pathway to deliver pro-apoptotic signals to the cancer cells, leading to the suppression of in vivo tumorigenesis. An extremely low concentration of cisplatin in addition to Ps-S-Oligos further up-regulated p53 activity, provoking massive apoptotic induction. These results suggest that the photodynamic antisense strategy has the great therapeutic potential in HPV-positive cervical cancers.     

Localization of ryanodine receptor 3 in the sinus endothelial cells of the rat spleen

The ultrastructural localization of ryanodine receptors (RyR) in sinus endothelial cells of the rat spleen was examined by confocal laser scanning and electron microscopy by using isoform-specific antibodies to each of the RyR isoforms. Immunofluorescence microscopy of tissue cryosections revealed RyR3 to be localized, with a strand-like form, in the superficial layer and within the cytoplasm of endothelial cells. Antibodies to RyR1 and RyR2 did not react indicating RyR3 was the predominant isoform. RyR3 was observed over the cortical layer of actin filaments in the apical part and beneath stress fibers in the basal part of the endothelial cells. The distribution of Ca²⁺-storing tubulovesicular-structures within endothelial cells was established by tissue sections treated with osmium ferricyanide selectively to stain the sarcoplasmic reticulum and transverse tubules in muscle cells; electron microscopy revealed densely stained tubulovesicular structures located throughout the sinus endothelial cells and interconnected at various sites. These structures closely apposed the plasma membrane at the apical, lateral, and basal surfaces of the cells and occasionally ran closely parallel to the plasma membrane and near to the mitochondria. Immunogold electron microscopy revealed RyR in the membranes of the nucleus, tubulovesicular structures, and subplasmalemmal cisternae. In the subplasmalemmal cisternae at the apical, lateral, and basal surfaces, RyR was detected on the membranes near to the plasma membrane. Labeling was also present on the membranes of tubulovesicular structures near to caveolae and on the cristae of the mitochondria. Thus, RyR probably participates in Ca²⁺ signal transduction and/or mechanosignal transduction in sinus endothelial cells.This work was supported by Grant-in-Aid for Scientific Research (C), Japan.     

Measurement of Individual Doses of Radiation by Personal Dosimeter Is Important for the Return of Residents from Evacuation Order Areas after Nuclear Disaster

To confirm the availability of individual dose evaluation for the return of residents after the accident at the Fukushima Dai-ichi Nuclear Power Plant (FNPP), we evaluated individual doses of radiation as measured by personal dosimeters in residents who temporarily stayed in Evacuation Order Areas in Kawauchi village, which is partially located within a 20 km radius of the FNPP. We also compared individual doses with the external radiation doses estimated from the ambient dose rates and with doses estimated from the concentrations of radionuclides in the soil around each individual’s house. Individual doses were significantly correlated with the ambient doses in front of the entrances to the houses (r = 0.90, p<0.01), in the backyards (r = 0.41, p<0.01) and in the nearby fields (r = 0.80, p<0.01). The maximum cumulative ambient doses in the backyards and fields around the houses were 6.38 and 9.27 mSv/y, respectively. The maximum cumulative individual dose was 3.28 mSv/y, and the median and minimum doses were 1.35 and 0.71 mSv/y. The estimated external effective doses from concentrations of artificial radionuclides in soil samples ranged from 0.03 to 23.42 mSv/y. The individual doses were moderately correlated with external effective doses in the backyards (r = 0.38, p<0.01) and in the fields (r = 0.36, p<0.01); however, the individual doses were not significantly correlated with the external effective doses in front of the entrances (r = 0.01, p = 0.92). Our study confirmed that individual doses are low levels even in the evacuation order area in Kawauchi village, and external effective dose levels are certainly decreasing due to the decay of artificial radionuclides and the decontamination of contaminated soil. Long-term follow-up of individual doses as well as internal-exposure doses, environmental monitoring and reconstruction of infrastructure are needed so that residents may return to their hometowns after a nuclear disaster.     

Cyclosporine A inhibits the secretion of certain anterior pituitary hormones in patients with nephrotic syndrome.

To clarify the effects of cyclosporine A (CsA) on the secretion of serum thyrotropin (TSH), prolactin (PRL), luteinizing hormone (LH) and follicular stimulating hormone (FSH), we performed TRH and LH-RH testing in 4 patients with the nephrotic syndrome before and after the administration of CsA, 6 mg/kg/day for 4 to 12 weeks. Prior to CsA all patients responded normally to TRH with respect to TSH and PRL secretion. Two patients showed normal response of LH and FSH to LH-RH stimulation while the response in 2 other patients, who were both menopausal, was exaggerated. By the third or fourth week of CsA administration the basal and peak TSH and PRL values declined significantly in all patients in response to TRH stimulation while those of LH and FSH showed only a modest decrease in response to LH-RH stimulation. Two to 4 weeks after the cessation of CsA the response of TSH, PRL and FSH returned to the pretreatment level. These observations suggest that: 1) CsA exerts an inhibitory effect on the secretion of at least TSH and PRL in humans, and 2) the effect of CsA on the pituitary may be partially reversible after the cessation of the therapy.     

Dynamics of blood coagulation factor XIII in ulcerative colitis and preliminary study of the factor XIII concentrate

Blood coagulation studies were performed in twenty patients with ulcerative colitis (UC). At the active stage of UC, a marked increase in platelet count and fibrinogen concentration, and a marked decrease in Factor XIII activity level were observed. At the active stage of UC, four patients were treated with Factor XIII concentrate leading to reduction of pain, bleeding and endoscopic findings.