Effects of the chelating agent DTPA on naturally accumulating metals in the body.

Diethylenetriaminepentaacetate (DTPA) is the most widely used chelating agent for Pu and Am. Volunteers were assigned to receive intravenous injections or aerosol inhalations of 1 g of DTPA on days 1-4; volunteers received once daily injections of CaDTPA or ZnDTPA, CaDTPA inhalation as an aerosol, or CaDTPA injection on day 1 and ZnDTPA on days 2-4. CaDTPA injection or inhalation increased the excretion rates of Zn in urine with concomitantly reduced levels of serum Zn. Injection of CaDTPA reduced activities of serum alkaline phosphatase (AP) in parallel with the kinetics of Zn, whereas CaDTPA and ZnDTPA injection reduced activities of lactate dehydrogenase (LDH), and reduced activities of creatinine kinase (CK) were observed upon CaDTPA injection and its inhalation. Intravenous administration of CaDTPA and ZnDTPA enhanced excretion rates of Mn in urine, whereas transient reduction of Mn levels in serum was detected only via CaDTPA injection. Both CaDTPA and ZnDTPA transiently reduced levels of Mg in serum without affecting the excretion rates. On the other hand, both DTPAs increased excretion rates of toxic metals such as Pb and Cd, and CaDTPA also increased the rates of Hg. These results suggest that DTPA, and especially CaDTPA, removes essential metals and that the activities of these metalloenzymes are good indicators for the imbalance of essential metals during the DTPA administration. Our results also show that CaDTPA injection is more potent for removing these metals than ZnDTPA and inhalation of CaDTPA, and DTPA may be useful for the treatment of acute heavy metal poisoning with Pb, Cd, or Hg.

Early Intake of Radiocesium by Residents Living Near the Tepco Fukushima Dai-ichi Nuclear Power Plant After the Accident. Part 2: Relationship Between Internal Dose and Evacuation Behavior in Individuals.

The Tokyo Electric Power Company's Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident exposed members of the public to radiation. This study analyses the relation between personal behavior data obtained from 112 out of 174 subjects who underwent whole-body measurements by the National Institute of Radiological Sciences (NIRS) during the period from 27 June to 28 July 2011 and their committed effective doses (CEDs) from Cs and Cs. The whereabouts of the 112 persons living in municipalities near the FDNPP (mainly, Namie town) on several days in March 2011 are graphed on maps. It was confirmed that most subjects started evacuation promptly and had left the 20-km-radius of the FDNPP by the end of 12 March. The individual CEDs were poorly correlated with the person's distances from the FDNPP at any day in March. Meanwhile, the percentage of persons remaining within the 20-km radius of the FDNPP was 100% at 16:00 on 12 March and 42.9% at 0:00 on 15 March for those with CEDs > 0.1 mSv, whereas the corresponding values were much lower for those with CEDs ≤ 0.1 mSv. This suggests that the time of evacuation would be one of the crucial factors for the early intake; however, more personal behavior data are needed to be analyzed to clarify the relevance to the individual internal dose.

Early Intake of Radiocesium by Residents Living Near the TEPCO Fukushima Dai-Ichi Nuclear Power Plant after the Accident. Part 1: Internal Doses Based on Whole-body Measurements by NIRS.

The Tokyo Electric Power Company's Fukushima Dai-ichi Nuclear Power Plant (FDNPP) accident in 2011 resulted in a release of radionuclides into the environment (I: 142.9 PBq, Cs:12.4 PBq). This study presents the results of internal doses to 174 residents living near the FDNPP at the time of the accident based on whole-body (WB) measurements performed by the National Institute of Radiological Sciences (NIRS) during the period between 27 June and 28 July 2011. The 174 subjects consisted of 125 adults (≥18-y) and 49 children (<18-y) and included 90 persons of Namie town, one of the municipalities heavily contaminated with the radionuclides. The number of subjects with significant detection of both Cs and Cs was relatively small: 28.8% for the adults and 4.1% for the children. A significant gender difference in the Cs detection rate (males > females) was observed in the adults but not the children. In this study, the committed effective dose (CED) from Cs and Cs was calculated based on individual WB contents (Cs) corrected against body size, the observed body content ratio of Cs to Cs, and the assumed intake scenario (namely, acute inhalation of Type F compounds on 12 March 2011 when the first explosive event occurred at the site of the FDNPP). The 90th-percentile CED value for the adults was around 0.1 mSv and the maximum CED (0.63 mSv) was found in an elderly male. Comparable CED results were obtained in other WB measurements subsequently performed by the Japan Atomic Energy Agency (JAEA) in a similar manner to that of the NIRS, suggesting that the contribution of ingestion to the WB content observed would be trivial for most of the JAEA subjects. The intake ratio of I to Cs was evaluated to be 3~5 based on the I thyroid measurement data of Tokonami et al. Using the average intake ratio of 3.8, the resulting median and maximum thyroid-equivalent doses to the adult subjects of this study were estimated at 3.5 mSv and 84 mSv, respectively.

Lessons Learned from the Accident at the Fukushima Dai-ichi Nuclear Power Plant-More than Basic Knowledge: Education and its Effects Improve the Preparedness and Response to Radiation Emergency.

A huge earthquake struck the northeast coast of the main island of Japan on 11 March 2011 triggering an extremely large tsunami to hit the area. The earthquake and tsunami caused serious damage to the Fukushima nuclear power plants (NPPs) of Tokyo Electric Power Company (TEPCO), resulting in large amounts of radioactive materials being released into the environment. The major nuclides released were (131)I, (134)Cs and (137)Cs. The deposition of these radioactive materials on land resulted in a high ambient dose of radiation around the NPPs, especially within a 20-km radius. Dose assessments based on behavior survey and ambient dose rates revealed that external doses to most residents were lower than 5 mSv, with the maximum dose being 25 mSv. It was fortunate that no workers from the NPPs required treatment from the viewpoint of deterministic effects of radiation. However, a lack of exact knowledge of radiation and its effects prevented the system for medical care and transportation of contaminated personnel from functioning. After the accident, demands or requests for training courses have been increasing. We have learned from the response to this disaster that basic knowledge of radiation and its effects is extremely important for not only professionals such as health care providers but also for other professionals including teachers.

The accident at the Fukushima Daiichi Nuclear Power Plant in 2011.

A huge earthquake struck the northeast coast of the main island of Japan on 11 March 2011, triggering a tsunami with more than 10-m-high waves hitting the area. The earthquake was followed by numerous sustained aftershocks. The earthquake and aftershocks left almost 16,000 people dead and more than 2,800 missing (as of 11 March 2014). The earthquake affected the Fukushima Daiichi Nuclear Power Plant (NPP) of Tokyo Electric Power Company (TEPCO), causing serious damage to the NPP and resulting in large amounts of radioactive materials being released into not only controlled areas but also the environment. Damage was caused to the cooling systems of the NPP, although they automatically shut down after the earthquake. The trouble with the cooling systems led to hydrogen explosions and core meltdown. The major nuclides released on land were ¹³¹I, ¹³4Cs, and ¹³7Cs. The release of these radioactive materials resulted in contamination of first responders and workers and also a high ambient dose of radiation around the NPP. The local hospital system, including that for radiation emergency medicine, was dysfunctional. Hospitals that had been designated as radiation emergency facilities were not able to function because the earthquake and tsunami had caused damage to their facilities; some of these were located within a 20-km radius of the NPP and in the evacuation areas. Local fire department personnel were also ordered to evacuate. Fukushima prefecture changed the screening level required for decontamination from 13,000 to 100,000 cpm, with decontamination by wiping being performed for over 13,000 cpm. However, as hospitals and fire departments had to abide by lower levels than that of the prefecture for receiving or transporting contaminated patients, these personnel could not accept or transport contaminated people from the NPPs. In addition, hospitals not designated as radiation emergency facilities would not receive patients from the NPPs because of concerns about the health effects of radiation. From this disaster, it was learned that basic knowledge of radiation and its effects is extremely important for health care providers.

Medical management of the consequences of the Fukushima nuclear power plant incident.

A huge earthquake struck the northeast coast of the main island of Japan on March 11, 2011, triggering a tsunami with 14-15 meter-high waves hitting the area. The earthquake was followed by numerous sustained aftershocks. The earthquake affected the nuclear power plant (NPP) in Fukushima prefecture, resulting in large amounts of radioactive materials being released into the environment. The major nuclides released on land were ¹³¹I, ¹³4Cs, and ¹³7Cs. Therefore, almost 170,000 people had to be evacuated or stay indoors. Besides the NPP and the telecommunications system, the earthquake also affected infrastructures such as the supplies of water and electricity as well as the radiation monitoring system. The local hospital system was dysfunctional; hospitals designated as radiation-emergency facilities were not able to function because of damage from the earthquake and tsunami, and some of them were located within a 20 km radius of the NPP, the designated evacuation zone. Local fire department personnel were also asked to evacuate. Furthermore, the affected hospitals had not established their evacuation plans at that time. We have learned from this "combined disaster" that the potential for damage to lifelines as well as the monitoring systems for radiation in case of an earthquake requires our intense focus and vigilance, and that hospitals need comprehensive plans for evacuation, including patients requiring life support equipment during and after a nuclear disaster. There is an urgent need for a "combined disaster" strategy, and this should be emphasized in current disaster planning and response.

[Radiation emergency medical preparedness in Japan--lessons learned from the Fukushima accident].

Although radiation exposure accidents fortunately occur only rarely, potential sources for exposure accidents can be found anywhere. When persons are accidentally exposed to radiation, physicians may be involved in their assessment and care; of course, their early diagnosis and dose assessment are crucial. After the criticality accident at Tokaimura in 1999, the system of radiation emergency medical preparedness has been further strengthened for nuclear facilities in Japan. In the revised system, hospitals involved were classified into three levels, depending on their locations and capabilities. The Great East Japan Earthquake attacked the Pacific coast area of eastern Japan on 11 March 2011. This earthquake and tsunami caused serious damage to the nuclear power plants of Tokyo Electric Power Co.(TEPCO) in Fukushima Prefecture; a large amount of radionuclides such as iodine and cesium were released into the environment. Since the revised system was focused on treatment of heavily exposed patients and knowledge on radiation was not enough for medical staff, many problems were raised at hospitals and fire departments in this disaster.

Exogenously-added copper/zinc superoxide dismutase rescues damage of endothelial cells from lethal irradiation.

The vascular endothelium is important for the early and late effects observed in lethally irradiated tissue and organs. We examined the effects of exogenously added superoxide dismutase on cell survival and angiogenesis in lethally irradiated human primary umbilical vein endothelial cells. Cell survival was significantly improved in superoxide dismutase-treated cells; the addition of superoxide dismutase to cells after irradiation was also effective for increased survival, as it was before irradiation. Moreover, treatment of cells with superoxide dismutase enhanced the phosphorylation of mitogen-activated protein/extracellular signal-regulated kinase/extracellular signal regulated kinases 1 and 2 in human primary umbilical vein endothelial cells. The addition of superoxide dismutase to cells after irradiation attenuated the reduction of angiogenesis by irradiation, and inhibition of the mitogen-activated protein/extracellular signal-regulated kinase/extracellular signal regulated kinases signaling pathway abrogated the rescue effect of superoxide dismutase. Our results suggest that superoxide dismutase rescues human primary umbilical vein endothelial cells from endothelial dysfunction caused by irradiation via a pathway requiring activation of mitogen-activated protein/extracellular signal-regulated kinase/extracellular signal regulated kinases 1 and 2.

Acceleration of regeneration of mucosa in small intestine damaged by ionizing radiation using anabolic steroids.

Damage to intestine is a serious problem after accidental radiation exposure. To examine substances to ameliorate damage by postirradiation administration, we focused on the regeneration process after irradiation of the intestine. Using experimental systems, the effects of clinically used sex hormones on regeneration were compared. An anabolic steroid, nandrolone (19-nortestosterone), stimulated proliferation in IEC-6 epithelial cells. A single injection of 19-nortestosterone ester with prolonged action into mice 24 h after abdominal irradiation at a lethal dose of 15.7 Gy showed significant life-saving effects. Regeneration indicators such as microcolonies of BrdU-incorporated cells at day 5 and c-myb mRNA expression levels at day 4 were enhanced by 19-nortestosterone administration. In contrast, high concentrations of estradiol inhibited growth of IEC-6 cells. Treatment of abdominally irradiated mice with estradiol ester decreased levels of regeneration indicators and survival. These results suggest the effectiveness of the anabolic steroid as well as the importance of manipulation of steroid receptors in the recovery of mucosa damaged by radiation.

Alterations in manganese, copper, and zinc contents, and intracellular status of the metal-containing superoxide dismutase in human mesothelioma cells.

BACKGROUND AND AIM: Molecular diagnostics and therapeutics of human mesothelioma using disease-related markers present major challenges in clinical practice. To identify biochemical alternations that would be markers of human mesothelioma, we measured the intracellular steady-state levels of biologically important trace metals such as manganese (Mn), copper (Cu), and zinc (Zn) in a human mesothelial cell line, MeT-5A, and in five human mesothelioma cell lines (MSTO-211H, NCI-H226, NCI-H2052, NCI-H2452, ACC-MESO-1) by inductively coupled plasma-mass spectrometry (ICP-MS). We also aimed to investigate whether the alterations were related to the intracellular status of metal-containing superoxide dismutase (SOD). RESULTS: There were no significant differences in the contents of the trace metals among MeT-5A, MSTO-211H, and ACC-MESO-1 cells. However, each of the other three mesothelioma cell lines had a unique characteristic in terms of the intracellular amounts of the metals; NCI-H226 contained an extremely high level of Mn, an amount 7.3-fold higher than that in MeT-5A. NCI-H2052 had significantly higher amounts of Cu (3.4-fold) and Zn (1.3-fold) compared with MeT-5A. NCI-H2452 contained about 5.8-fold the amount of Cu and 2.5-fold that of Mn compared with MeT-5A. As for the intracellular levels of copper/zinc-SOD (Cu/Zn-SOD) and manganese-SOD (Mn-SOD), those of Cu/Zn-SOD were relatively unchanged among the cells tested, and no notable correlation with Cu or Zn contents was observed. On the other hand, all mesothelioma cells highly expressed Mn-SOD compared with MeT-5A, and a very high expression of the enzyme with a robust activity was observed in the two mesothelioma cells (NCI-H226, NCI-H2452) containing a large amount of Mn. CONCLUSIONS: In comparison with MeT-5A human mesothelial cells, some human mesothelioma cells had significantly higher amounts of Mn or Cu and one mesothelioma cell had a significantly higher amount of Zn. Interestingly, all mesothelioma cells overexpressed Mn-SOD compared with MeT-5A, and the cells whose Mn-SOD activity was increased contained higher amounts of Mn. It seemed that intracellular Mn content was positively correlated with Mn-SOD, suggesting that the intracellular Mn level is associated with Mn-SOD activity. These biochemical signatures could be potential disease-related markers of mesothelioma.

Catalase regulates cell growth in HL60 human promyelocytic cells: evidence for growth regulation by H(2)O(2).

Reactive oxygen species (ROS) including hydrogen peroxide (H(2)O(2)) are generated constitutively in mammalian cells. Because of its relatively long life and high permeability across membranes, H(2)O(2) is thought to be an important second messenger. Generation of H(2)O(2) is increased in response to external insults, including radiation. Catalase is located at the peroxisome and scavenges H(2)O(2). In this study, we investigated the role of catalase in cell growth using the H(2)O(2)-resistant variant HP100-1 of human promyelocytic HL60 cells. HP100-1 cells had an almost 10-fold higher activity of catalase than HL60 cells without differences in levels of glutathione peroxidase, manganese superoxide dismutase (MnSOD), and copper-zinc SOD (CuZnSOD). HP100-1 cells had higher proliferative activity than HL60 cells. Treatment with catalase or the introduction of catalase cDNA into HL60 cells stimulated cell growth. Exposure of HP100-1 cells to a catalase inhibitor resulted in suppression of cell growth with concomitant increased levels of intracellular H(2)O(2). Moreover, exogenously added H(2)O(2) or depletion of glutathione suppressed cell growth in HL60 cells. Extracellular signal regulated kinase 1/2 (ERK1/2) was constitutively phosphorylated in HP100-1 cells but not in HL60 cells. Inhibition of the ERK1/2 pathway suppressed the growth of HP100-1 cells, but inhibition of p38 mitogen-activated protein kinase (p38MAPK) did not affect growth. Moreover, inhibition of catalase blocked the phosphorylation of ERK1/2 but not of p38MAPK in HP100-1 cells. Thus our results suggest that catalase activates the growth of HL60 cells through dismutation of H(2)O(2), leading to activation of the ERK1/2 pathway; H(2)O(2) is an important regulator of growth in HL60 cells.

Endogenous production of TNFalpha is a potent trigger of NFkappaB activation by irradiation in human monocytic cells THP-1.

Irradiation causes DNA damage and induces neoplastic transformation. In response to irradiation, cells induce genes or activate proteins that protect themselves from the external insult. Nuclear factor kappaB (NFkappaB) activates transcription of target genes and plays important roles in inflammation. We studied the mechanism(s) for activation of NFkappaB by irradiation in human monocytic cells THP-1. Gel mobility shift assays showed that irradiation stimulated the NFkappaB-DNA binding activity of nuclear extracts from these cells. Western blot analysis using polyclonal antibody against phosphorylated IkappaB protein showed that irradiation increased the levels of phosphorylated IkappaB. The production of tumor necrosis factor alpha (TNFalpha) was stimulated by irradiation in these cells. Treatment with exogenously added TNFalpha also stimulated the NFkappaB binding activity with concomitant degradation of IkappaB. Further study found that the activation of NFkappaB by irradiation was inhibited by a neutralizing anti-TNFalpha antibody. Macrophages from TNFalpha-deficient mice were also defective in the irradiation-induced activation of NFkappaB. These results indicate that endogenous production of TNFalpha in macrophages/monocytes is required for NFkappaB activation by irradiation. Our data also suggest that TNFalpha in monocytes/macrophages exposed to irradiation is involved in signal transduction network initiation.

An accidental exposure by a medical linear accelerator under construction.

A radiation accident occurred at a medical linear accelerator facility under construction in Japan. The radiation source was a 3- and 6-MV potential drop accelerator designed to produce X-rays for radiation therapy. This accelerator was also capable of producing a 5 to14-MV swept electron beam. During setting up, an operator turned on the accelerator to test the beam not knowing that a man was working on the ceiling above the accelerator. Thus, an X-ray beam was emitted against the ceiling and the man was exposed to 10-MV of X-ray irradiation. However, no obvious physical symptoms were noted. Dose estimation was made from reconstruction of the accident and clinical examinations including chromosome analysis. Mean dose of the whole body ranged from 70 to 180 mSv. Estimated dose from his right foot to hand was between180 to 900 mSv.

Role of reactive oxygen species in cells overexpressing manganese superoxide dismutase: mechanism for induction of radioresistance.

Manganese superoxide dismutase (MnSOD) catalyzes the dismutation of superoxide anions (O(2)(-)) into hydrogen peroxide (H(2)O(2)). We altered the intracellular status of reactive oxygen species by introducing human MnSOD cDNA into the human ovarian cancer cell line SK-OV-3. The overexpression of MnSOD inhibited cell growth and induced a concomitant increase in the level of H(2)O(2) in SK-OV-3 cells. The cells overexpressing MnSOD were more resistant to irradiation than parental cells. MnSOD overexpression shortened the G(2)-M duration in irradiated cells. Either inhibition of p38 mitogen-activated protein kinase (p38MAPK) or scavenging free radicals blocked the induction of radioresistance by MnSOD and also abolished the shortening of the G(2)-M duration with concomitant inhibition of p38MAPK phosphorylation. Irradiation increased the generation of H(2)O(2) even more in these transfectants. These results suggest that the accumulated H(2)O(2) potentiated the activation of p38MAPK after irradiation in cells overexpressing MnSOD, which led to the protection of cells from irradiation-mediated cell death through the G(2)-M checkpoint. SK-OV-3 cells had no constitutive expression of p53, and the overexpression of MnSOD and/or irradiation did not induce p53 or p21(WAF1), which causes cell cycle arrest. Thus, our results suggest that MnSOD alters the cell cycle progression of irradiated cells independently of p53 and p21(WAF1).

Hsp25 regulates the expression of p21(Waf1/Cip1/Sdi1) through multiple mechanisms.

Exposure of cells to external stresses leads to the induction or activation of certain proteins. Expression of heat shock proteins (Hsps) is induced in response to these stresses. Hsps are known to have molecular chaperone activities; but recent studies have shown that Hsps have a variety of functions such as the triggering of proliferation, differentiation, and apoptosis of cells. Previously, we found that overexpression of a 25 kDa Hsp (Hsp25) induced expression of cell cycle inhibitory protein p21 (Waf1/Cip1/Sdi1) in murine fibroblastoid L929 cells. However, the mechanisms underlying the induction of p21 by Hsp25 are unknown. In the present study, we investigated the mechanisms underlying the regulation of p21 expression by Hsp25 in these cells. The introduction of Hsp25 cDNA stimulated the accumulation of p21 transcripts through transcriptional but not posttranscriptional regulation in these cells. We also found that overexpression of Hsp25 markedly increased the translational rate of p21 and stabilized the protein. Studies involving proteasome inhibitors and Western blot analysis for ubiquitination of p21 demonstrated that the stabilization of p21 is regulated through a ubiquitin-independent pathway. However, no direct association of Hsp25 with p21 was observed. These findings suggest that Hsp25 induces p21 expression through multiple mechanisms, and that transcriptional, translational, and post-translational regulation are important in the regulation of p21.