Satoru Monzen

The time variation of dose rate artificially increased by the Fukushima nuclear crisis

A car-borne survey for dose rate in air was carried out in March and April 2011 along an expressway passing northwest of the Fukushima Dai-ichi Nuclear Power Station which released radionuclides starting after the Great East Japan Earthquake on March 11, 2011, and in an area closer to the Fukushima NPS which is known to have been strongly affected. Dose rates along the expressway, i.e. relatively far from the power station were higher after than before March 11, in some places by several orders of magnitude, implying that there were some additional releases from Fukushima NPS. The maximum dose rate in air within the high level contamination area was 36 μGy h−1, and the estimated maximum cumulative external dose for evacuees who came from Namie Town to evacuation sites (e.g. Fukushima, Koriyama and Nihonmatsu Cities) was 68 mSv. The evacuation is justified from the viewpoint of radiation protection.

Activity concentrations of environmental samples collected in Fukushima Prefecture immediately after the Fukushima nuclear accident

Radionuclide concentrations in environmental samples such as surface soils, plants and water were evaluated by high purity germanium detector measurements. The contribution rate of short half-life radionuclides such as 132I to the exposure dose to residents was discussed from the measured values. The highest values of the 131I/137Cs activity ratio ranged from 49 to 70 in the environmental samples collected at Iwaki City which is located to the south of the F1-NPS. On the other hand, the 132I/131I activity ratio in the same environmental samples had the lowest values, ranging from 0.01 to 0.02. By assuming that the 132I/131I activity ratio in the atmosphere was equal to the ratio in the environmental samples, the percent contribution to the thyroid equivalent dose by 132I was estimated to be less than 2%. Moreover, the contribution to the thyroid exposure by 132I might be negligible if 132I contamination was restricted to Iwaki City.

Relationship between Radiosensitivity and Nrf2 Target Gene Expression in Human Hematopoietic Stem Cells

Abstract NFE2-related factor 2 (Nrf2), which belongs to the cap “n” collar family of basic region leucine zipper transcription factors, is a key protein in the coordinated transcriptional induction of expression of various antioxidant genes. The purpose of this study was to analyze the expression of Nrf2 target genes, such as heme oxygenase 1 (HO-1), ferritin heavy polypeptide 1 (FTH1), NAD(P)H dehydrogenase, quinone 1 (NQO1), glutamate-cysteine ligase catalytic subunit, glutamate-cysteine ligase modifier subunit, glutathione reductase (GSR) and thioredoxin reductase 1 (TXNRD1), after X irradiation of CD34+ cells that were prepared from human placental/umbilical cord blood hematopoietic stem cells (HSCs). We evaluated the relationship between radiosensitivity and expression of Nrf2 target genes in HSCs. The number of colony-forming cells derived from 2-Gy-irradiated HSCs decreased to approximately 20% of the nonirradiated control. At the same time, the mRNA expression of HO-1, FTH1, NQO1, GSR and TXNRD1 was significantly increased after X irradiation. A statistically significant negative correlation was observed between the surviving fraction of HSCs and the intrinsic NQO1 mRNA expression, indicating that HSCs in which NQO1 mRNA levels are low may also be radioresistant. The present results suggest that the antioxidant system associated with Nrf2 is involved in the radiosensitivity of HSCs.

Individual Radiation Exposure Dose Due to Support Activities at Safe Shelters in Fukushima Prefecture

Immediately after the accidents in the nuclear power stations in Fukushima on March 11, the Japanese Government ordered the evacuation of the residents within a 20-km radius from the station on March 12, and asked various institutions to monitor the contamination levels of the residents. Hirosaki University, which is located 355 km north of Fukushima City, decided to send support staff to Fukushima. This report summarizes the results of the exposure of 13 individual teams from March 15 to June 20. The support teams surveyed more than 5,000 people during this period. Almost all subjects had external contamination levels of less than 13 kcpm on Geiger-Müller (GM) survey meter, which is categorized as “no contamination level.” The 1st team showed the highest external exposure dose, but the 4th team onward showed no significant change. Subsequently, the internal radiation exposure was measured using a whole body counter that indicated undetectable levels in all staff members. Although the measured external radiation exposure dose cannot have serious biological effects on the health of an individual, a follow-up study of the residents in Fukushima and other regions where the radioactive material has spread will be required for a long time.

Correlations of Cell Surface Antigens with Individual Differences in Radiosensitivity in Human Hematopoietic Stem/Progenitor Cells

Abstract To characterize the differences in the radiosensitivity of individual populations of human hematopoietic stem/progenitor cells (HSPCs), we examined the relationship among cell surface antigens, clonogenic potential and radiation survival. The expressions of CD34, CD38, CD45RA, CD110 and Tie-2, early differentiation pathway-related antigens in hematopoiesis, were analyzed on the surface of HSPCs enriched for CD34 antigen expression in 20 samples prepared from human placental/umbilical cord blood. A significantly positive relationship was observed between CD38 antigen and CD45RA and between CD110 and Tie-2. No significant relationship was observed in most cases among the antigens and the number of colony-forming cells (CFCs); however, the number of megakaryocytic progenitor cells correlated negatively with the percentage of Tie-2+ cells. The percentage Tie-2 cells correlated significantly with the surviving fraction of CFCs irradiated with 2 Gy of X rays, suggesting that the radiosensitivity of individual CFC populations is related to the percentage of Tie-2-expressing cells. In addition, the number of progenitor cells closely correlated with the surviving fraction after 2 Gy of X rays. These results suggest that the radiosensitivity of individual HSPC populations is related to the number of progenitor cells in the population especially dependent on the presence of immature HSPCs such as Tie-2+ cells.

Ex vivo expansions of megakaryocytopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free culture supplemented with proteoglycans extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of whale.

As a possible approach to the treatment of thrombopocytopenia, the ex vivo expansion of megakaryocytic progenitor cells may be a useful tool to accelerate platelet recovery in vivo. Our objective was to assess the promoting effect of proteoglycans in a serum-free culture condition using human cord blood CD34(+) cells. Highly purified proteoglycan (PG) extracted from the nasal cartilage of salmon heads and the nasal septum cartilage of a whale were applied to the ex vivo expansion of megakaryocytopoiesis and thrombopoiesis from placental and umbilical cord blood CD34(+) cells in serum-free cultures stimulated with a combination of thrombopoietin (TPO) and interleukin-3 (IL-3). Each PG (0.5 and 5 mug) was applied to the culture with three different concentrations of TPO (50, 5 and 0.5 ng/ml) and IL-3 (100, 10 and 1 ng/ml). Both of the PGs showed no promoting effects on the mononuclear cell proliferation rate in any of the cultures. However, the whale-PG promoted the generation of megakaryocytic progenitor cells and megakaryocytes in the culture with a lower dose of cytokines, respectively. In addition, whale-PG led to a significant increase in CD42a(+) particles which seemed to be platelets. While the salmon-PG failed to promote such production in almost all of the cultures. Although whale-PG is an attractive molecule for the ex vivo expansion of human megakaryocytopoiesis, its action may depend on the glycosaminoglycans sulfation pattern and the ability of the binding affinity and the kinetics to interact with the cytokines and hematopoietic stem/progenitor cells.

Severe Damage of Human Megakaryocytopoiesis and Thrombopoiesis by Heavy-Ion Beam Radiation

Abstract Takahashi, K., Monzen, S., Eguchi-Kasai, K., Abe, Y. and Kashiwakura, I. Severe Damage of Human Megakaryocytopoiesis and Thrombopoiesis by Heavy-Ion Beam Radiation. Radiat. Res. 168, 545–551 (2007). Heavy ions have a unique efficacy for tumor control in radiotherapy. To clarify the effects of heavy-ion beams on hematopoietic stem/progenitor cells, the effects of carbon-ion beams on megakaryocytopoiesis and thrombopoiesis in CD34+ cells derived from human placental and umbilical cord blood were investigated. The cells were exposed to carbon-ion beams (LET = 50 keV/μm) and then were treated with thrombopoietin (TPO) alone or TPO plus other cytokines. Megakaryocytic progenitor cells, such as megakaryocyte colony-forming units (CFU-Meg), were far more sensitive to carbon-ion beams than to X rays, and no restoration of carbon-ion beam-irradiated CFU-Meg by treatment with any cytokine combination was observed. However, total cell expansion in liquid culture was not different after either carbon-ion beam or X irradiation of CD34+ cells. The activation of γ-H2AX, a marker of DNA double strand-breaks (DSBs), was promoted by the cytokine treatment in X-irradiated CD34+ cells but not in carbon-ion-irradiated cells. These results showed that carbon-ion beams inflicted severe damage on megakaryocytopoiesis and thrombopoiesis and that a better combination of cytokines and other agents may be needed to stimulate the recovery of hematopoietic cells and repair this damage.

Exosomes derived from SW480 colorectal cancer cells promote cell migration in HepG2 hepatocellular cancer cells via the mitogen-activated protein kinase pathway.

Exosomes are membrane-derived extracellular vesicles that have recently been recognized as important mediators of intercellular communication. In the present study, we investigated the effects of exosomes derived from SW480 colorectal cancer cells in recipient HepG2 hepatocellular cancer cells. We demonstrated that SW480-derived exosomes were taken up by the recipient HepG2 cells via dynamin-dependent endocytosis and were localized to the HepG2 lysosomes. In addition, SW480-derived exosomes induced the phosphorylation of extracellular signal-regulated kinase (ERK)1/2 following their uptake into HepG2 cells. Of note, these changes occurred during the early phase after exosome treatment. Furthermore, SW480-derived exosomes promoted the migration of recipient HepG2 cells in a wound-healing assay, which was suppressed by pretreatment with U0126, an upstream inhibitor of ERK1/2. These results indicated that SW480-derived exosomes activated a classical mitogen-activated protein kinase pathway in recipient HepG2 cells via dynamin-dependent endocytosis and subsequently enhanced cell migration by ERK1/2 activation. Our results provide new insights into the regulation of cellular functions by exosomes.

Radiation sensitivities in the terminal stages of megakaryocytic maturation and platelet production.

Abstract These studies examined the effects of X radiation and interleukin 3 (IL-3), which is an effective cytokine for the generation of megakaryocytopoiesis from X-irradiated hematopoietic stem/progenitor cells, on the terminal process of human megakaryocytopoiesis and thrombopoiesis. Mature megakaryocytes were induced by culturing CD34+ cells from normal human peripheral blood in a serum-free liquid culture stimulated with thrombopoietin. The experiments contained the following groups: control cultures with nonirradiated cells incubated for 15 days; cultures treated with IL-3 on day 7 or day 11, cultures irradiated with 2 Gy on day 7 or day 11, and cultures treated with IL-3 immediately after X irradiation. The nonirradiated control cultures produced megakaryocytes from day 7, and both the megakaryocyte and platelet generation reached a peak on day 12–13. When X irradiation was performed on day 7, both the megakaryocyte and platelet numbers decreased remarkably, while no significant effect was observed on those numbers when cultures were X-irradiated on day 11. IL-3 showed neither protective nor promoting effects on the terminal stages of megakaryocytic maturation and platelet production. The results demonstrated that mature megakaryocytes are radiosensitive but that the radiosensitivity decreased with the terminal stages of megakaryocytic maturation, especially for the megakaryocytes entering into proplatelet formation.

Megakaryocytopoiesis and Thrombopoiesis in Hematopoietic Stem Cells Exposed to Ionizing Radiation

Hematopoietic processes, especially megakaryocytopoiesis and thrombopoiesis, are highly sensitive to extracellular oxidative stresses such as ionizing radiation and chemotherapeutic agents. This study examined the terminal maturation of megakaryocytes and platelet production in hematopoietic stem/progenitor cells (HSPCs) exposed to ionizing radiation. Highly purified CD34+ cells derived from human placental/umbilical cord blood were exposed to X rays (2 Gy, 150 kVp, 20 mA; 0.5-mm aluminum and 0.3-mm copper filters) at a dose rate of approximately 1 Gy/min and then cultured in a serum-free medium supplemented with thrombopoietin and interleukin-3. The number of cells generated from X-irradiated CD34+ cells decreased with the time in culture. However, the fraction of CD34+Tie-2+ and CD41+Tie-2+ cells among the total cells generated from X-irradiated cells increased significantly in comparison to nonirradiated controls on day 7. In addition, the CD42a+ particles, which appeared to be platelets, generated from the X-irradiated HSPCs appeared to be normal. Quantitative real-time reverse transcriptase-polymerase chain reaction analysis of the expression of various genes in cells harvested from the cultures showed that the early hematopoiesis-related genes FLI1, HOXB4 and Tie-2, the cytokine receptor genes KIT and IL3RA, and the oxidative stress-related genes HO1 and NQO1 were upregulated on day 7. These results suggest that normal terminal maturation of megakaryocytes and platelet production occur in residual HSPCs after exposure to ionizing radiation despite the adverse effect of radiation on proliferation and differentiation of HSPCs. Ionizing radiation may have the potential to promote both megakaryocytopoiesis and thrombopoiesis.