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論文

Irradiation effect of gamma rays on cyanate ester/epoxy resins, 3

出崎 亮; 上地 宏樹*; 羽倉 吉彦*; 岸 肇*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 29, 2015/03

本研究では、J-PARCやITERに設置される超伝導磁石システム用電気絶縁材料として応用が期待されているシアネートエステル/エポキシ樹脂について、物理化学的特性変化の面から放射線照射効果を解明することを目的としている。これまでに、室温真空中において100MGyまでガンマ線を照射すると、水素, 一酸化炭素, 二酸化炭素が発生すること、樹脂のガラス転移温度が初期値の80%である180$$^{circ}$$Cまで低下することを明らかにしているが、本報では、放射線照射によって分解する化学構造の解明を核磁気共鳴法による分子構造解析により行なった。その結果、シアネートエステルとエポキシの間の架橋構造を構成するシアヌレート, イソシアヌレート, オキサゾリジノン骨格に結合するエーテル鎖が切断されていることが分かった。以上のことから、シアネートエステル/エポキシ樹脂に放射線を照射すると、シアネートエステルとエポキシの間で形成される架橋構造におけるエーテル鎖が切断され、ガス発生やガラス転移温度の低下が起こることが明らかになった。

論文

Complexation of lutetium-177 with bifunctional chelators in the presence of Mg, Cu, and Yb

渡辺 智; 橋本 和幸; 石岡 典子

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 102, 2015/03

As part of basic studies for the production of $$^{177}$$Lu capable of labeling of $$^{177}$$Lu-1,4,7,10-tetraazacyclododecan-N,N',N'',N'''-tetraacetic acid (DOTA)-antibody, $$^{177}$$Lu complexation of DOTA and diethylenetriamine-N,N,N',N'',N''-pentaacetic acid (DTPA) was investigated in the presence of competing metals: Mg, Cu, and Yb in addition to Ca, Fe, and Zn. The inhibition of $$^{177}$$Lu complexation by the competing metals was in the order of Mg, Ca $$<$$ Fe $$<$$ Yb $$<$$ Cu, Zn and Mg, Ca $$<$$ Zn $$<$$ Fe $$<$$ Cu $$<$$ Yb with DOTA and DTPA, respectively. Consequently, the elimination of Mg and Ca from the $$^{177}$$Lu final solution produced was not found to be necessary, while the elimination of Cu, Fe, Zn and Yb from the $$^{177}$$Lu final solution was found to be necessary. On the basis of the knowledge obtained, the method of elimination of the competing metals will be developed.

論文

Development of a scintillator for single-ion detection

横山 彰人; 加田 渉*; 佐藤 隆博; 江夏 昌志; 山本 春也; 神谷 富裕; 横田 渉

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 168, 2015/03

サイクロトロンでは、数百MeV以上の重イオンマイクロビームを用いたシングルイオンヒット技術が生物細胞などの照射実験で利用されており、高精度位置検出が必要とされている。しかし、現状の固体飛跡検出器では 照射後の薬品による表面処理と顕微鏡での照射位置の観察に時間を要し、リアルタイムでの検出が困難であった。そこで、ビームモニタに利用されているZnS等の発光体などを試用したが発光強度や位置分解能が十分ではなかった。本研究では、単結晶サファイアに賦活剤として注入するEuイオンの量や熱処理条件等の調整により、強いイオンルミネッセンス(Ion Luminescence: IL)を発する試料の開発と発光検出装置の感度試験を行った。予備実験として調製試料の表面にレーザー照射することにより、電子励起過程を伴って発光するフォトルミネッセンス(Photoluminescence: PL)を測定した結果、表面から30nm$$sim$$70nmにEuを7.5ions/nm$$^{3}$$を注入した後に、600$$^{circ}$$Cで30分間熱処理した試料のPL強度が最大だった。また、同試料を用いた発光検出装置の感度試験では、260MeV Neイオンを1秒間に200個のフルエンス率で1点照射することによってILを捕えることができた。しかしながら、シングルイオンヒット実験では、ビーム電流量が数cps程度と低いことから、感度の向上のために賦活剤の濃度の調整などの改良を今後行う。

論文

Dose responses of irradiated fresh papaya recorded on ESR at different temperature

菊地 正博; 鵜飼 光子*; 小林 泰彦

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 89, 2015/03

Electron spin resonance spectroscopy (ESR) has been utilized as a detection method of irradiated foods. However, there is a problem on ESR measurement for the water-rich foods. We have examined two techniques to solve the water problem. In two ESR detection methods for fresh fruits, dose response curves of signal intensities were obtained independently. Three responses show the direct proportional relationship, except for main peak signals of freeze-dried powders. In this study, we examined whether $$gamma$$-induced radicals of papaya fleshes are changed after different sample preparations. The right side peak intensity is good agreement with the estimation. $$gamma$$-induced radicals of the side peak are not affected by the sample preparations. This study indicates that the ESR detection methods for fresh fruits are reliable as a quantitative measurement.

論文

Analysis of radicals induced in irradiated amino acid using Pulse-ESR

岸田 敬吾*; 菊地 正博; 中村 秀夫*; 小林 泰彦; 鵜飼 光子*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 90, 2015/03

Radiation induced radicals in irradiated foods were analyzed with Pulse-ESR. The specimens were rice and wheat flour. We also analyzed 22 kinds of amino acids contained in foods to compare with the food specimens. Radiation dose level was 50,100 and 200 kGy. We could not detect any signals in all specimens before irradiation treatment. Upon radiation treatment one broad signal was detected. Using Pulse-ESR, we succeeded to detect field swept echo signal in some irradiated amino acid specimens and to analyze the relaxation times (T$$_{1}$$ and T$$_{2}$$). It is concluded that ESR is very useful to analyze the radiation induced radicals in irradiated foods.

論文

Metabolome analysis of leaves of ${it Rumex obtusifolius}$ L. irradiated with ion beams

宮城 敦子*; 北野 沙也佳*; 大野 豊; 長谷 純宏; 川合 真紀*; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 114, 2015/03

${it Rumex obtusifolius}$ (Polygonaceae) is a weed, which spreads all over the world. It contains amino acids and ascorbate (Vitamin C) more than other edible ${it Rumex}$ species (Miyagi et al., 2010). On the other hand, it highly accumulates soluble oxalate, which causes renal syndrome for vertebrates. Thus we aimed to lower oxalate contents in R. obtusifolius that can be used as a pasture by radiation breeding. In the present study, plant seeds irradiated with several doses of carbon ion beams ($$^{12}$$C$$^{6+}$$) were used to analyze effects of the radiation to oxalate contents and other metabolites using capillary electrophoresis-mass spectrometry (CE-MS). Results showed that the oxalate contents of leaves were increased by the irradiation of ion beams on the contrary to the case of the irradiation with $$gamma$$ rays. High oxalate plants tended to contain high levels of organic acids. Amino acids and phosphorylated organic acids were decreased in these plants.

論文

Screening of salt tolerant mutants regenerated from irradiated stem segments of Lombardy poplar (${it Populus nigra}$)

Biswas, K. K.*; 長谷 純宏; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 113, 2015/03

Species of the genus ${it Populus}$ are ecologically important trees and often serving as a vegetative pioneer species because of their fast growth rates. The rapid growth of poplar is dependent on access to sufficient water in plantation sites and this is problematic. To overcome this problem, a forward genetics approach has been taken for developing salt/drought tolerant mutants by combination of radiation mutagenesis and in vitro regeneration in ${it Populus}$. Previously we screened regenerated ${it Populus nigra}$ shoots regenerated from stems exposed with cobalt-60 source of $$gamma$$ rays and isolated two mutant candidates named ${it g1}$ and ${it g5}$. Only about 25% stem segments of both lines produced roots in RIM with 75 mM NaCl. The low frequency in root production in NaCl medium may be attributed to that these plants are chimeric. We also successfully obtained at least 2 salt tolerant candidates regenerated from stem segments irradiated with 320 MeV $$^{12}$$C$$^{6+}$$ ion beams.

論文

Development of new ${it Chrysanthemum Morifolium}$ pink mutants through ion beam irradiation

Zaiton, A.*; Affrida, A. H.*; Shakinah, S.*; Nurul Hidayah, M.*; 長谷 純宏; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 108, 2015/03

Developing new varieties of chrysanthemums is one of the emphases in Malaysian floriculture research as chrysanthemum is recognized as among the most important cut flower export for Malaysia. A combination of ion beam irradiation and ${it in vitro}$ organ culture techniques is very efficient in generating new ornamental varieties with novel traits and wider mutation spectrum. Therefore, in this project, ray floret cultures of pink chrysanthemum variety were irradiated at various doses with 320 MeV $$^{12}$$C$$^{6+}$$ ions, regenerated into plantlets and screened for morphological variations with an ultimate aim to produce mutants with useful horticultural traits. A series of screening was carried out at Nuclear Malaysia's glasshouse and MARDI Cameron Highlands. The mutants were selected and characterized morphologically. Based on these screenings, three mutants were identified based on their distinct flower and/or plant characteristics.

論文

ウンシュウミカンにおけるイオンビーム照射用試料調整方法の検討

松尾 洋一*; 長谷 純宏; 竹下 大樹*; 納富 麻子*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 111, 2015/03

中生ウンシュウミカンは、多胚性種子のために交雑育種による品種改良には多数の制約がある。このため新品種の開発は、雑種実生による品種改良は少なく、枝変わりもしくは珠心胚実生の中から突然変異個体を選抜している。通常の珠心胚実生の選抜試験では、特定の形質を得ることは困難であるため、イオンビーム照射を実施した珠心胚実生より突然変異個体を効率的に誘発するために、品種毎に試料の前処理方法を検討した。前処理方法として、実生の胚軸及び根部を切断してプラ壺へ移植する方法、ならびに組織の切断部位を少なくするために胚軸部のみを切断し、スチロールケースに配置して根部は水分を含ませた支持体で固定する方法の2通りを試験した。イオンビーム照射後の再分化率を比較した結果、前者の方法の方が再分化率が高いことがわかった。

論文

Molecular analysis of heavy ion induced mutations in budding yeast ${it S. cerevisiae}$

松尾 陽一郎*; 泉 佳伸*; 長谷 純宏; 坂本 綾子; 清水 喜久雄*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 119, 2015/03

高LETのイオンビームによる突然変異誘発の分子機構を明らかにするために、出芽酵母(${it Saccharomyces cerevisiae}$)を材料として研究を行ってきた。これまでに、LETの増加に伴って致死率および${it URA3}$遺伝子の突然変異率が上昇する傾向があること、ならびに突然変異の分布としてヌクレオソーム構造のリンカーDNAにおいて局所的に突然変異が誘発されることを示唆する結果を報告した。ここでは、突然変異が誘発される部位がヌクレオソーム構造に関係があるという仮説を検証するために、野生株と異なるヌクレオソーム構造の${it URA3}$遺伝子を持つ組換え株を作成し、突然変異誘発スペクトルを分析した。野生株での突然変異の位置と、ヌクレオソーム構造が異なる株での${it URA3}$遺伝子の突然変異位置には相違があり、突然変異が生じる位置がヌクレオソーム構造に依存することが示唆された。

論文

イオンビーム照射によるスプレーギクの花色変異の育種

田之頭 優樹*; 永吉 実孝*; 遠嶋 太志*; 長谷 純宏

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 109, 2015/03

本研究は、スプレーギク品種へのイオンビーム照射による花色ファミリー品種の効率的な育成方法に関するものである。夏秋スプレーギク品種「サザンチェルシー(桃色)」の葉片培養物へイオンビームを照射して得た変異集団からは、アントシアニンが減少した薄桃色及び白色、アントシアニンが増加した濃桃色、カロテノイドが増加した赤樺色及び黄樺色が得られた。育種目標とした白色の花色変異率は0.6%、赤樺色は1.4%であったが、黄色は0%であった。黄色の変異体を得るためには、アントシアニンが消失する変異と、カロテノイドが増加する2つの変異が必要と考えられるが、この2つの変異が独立して生じると仮定すると、1回の変異誘発で黄色変異体が得られる頻度は0.0084%と極めて低い。この結果から、白色変異株への再照射によってカロテノイドが増加した変異を獲得することが黄色変異体を獲得する上で高効率であると考えられた。

論文

Synthesis and ${{it in vitro}}$ evaluation of $$^{64}$$Cu-labeled peptide for tumor imaging

須郷 由美; 佐々木 一郎; 渡辺 茂樹; 大島 康宏; 石岡 典子

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 100, 2015/03

MARSGL peptide (H-Met-Ala-Arg-Ser-Gly-Leu-OH) has high affinity to the human epidermal growth factor receptor 2 (HER2) overexpressing in various tumor cells. Copper-64 ($$^{64}$$Cu) is a useful radionuclide in nuclear medicine, and can be produced by the cyclotron. In this study, we designed and synthesized $$^{64}$$Cu-labeled MARSGL peptide conjugated with 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid (DOTA) as a novel positron emission tomography (PET) imaging probe for HER2 overexpressing tumors. The formation of $$^{64}$$Cu-DOTA-MARSGL was determined by TLC and HPLC compared with a non-radioactive preparation. It was confirmed that $$^{64}$$Cu-DOTA-MARSGL was obtained in high radiochemical yield more than 94%. We also examined a stability of $$^{64}$$Cu-DOTA-MARSGL ${it in vitro}$. The chromatogram was not changed after incubation in physiological saline at 37$$^{circ}$$C overnight. In order to evaluate the usefulness as a PET imaging probe, further ${it in vitro}$ studies on the stability in human or mice plasma and the cellular uptake are in progress.

論文

Development of cesium-accumulating bacteria by ion beam breeding technology

佐藤 勝也; 上田 涼史郎; 長谷 純宏; 鳴海 一成*; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 117, 2015/03

Bioremediation uses biological organisms to solve an environmental problem. However, microorganisms are sometimes not enough effective to achieve efficient recovery. Therefore, enhancing the functionality of microorganisms is needed to promote the active use of bioremediation. This study began with the purpose of development of Cs-accumulating bacteria by ion beam breeding technology. We investigated which members are better suited for Cs-accumulation in the genus ${it Deinococcus}$ (${it D. radiodurans}$, ${it D. grandis}$, ${it D. proteolyticus}$, ${it D. radiopugnans}$, ${it D. geothermalis}$ and ${it D. murrayi}$) exhibit extraordinary radioresistant. The Cs concentration in the ${it Deinococcus}$ cells exhibited over 4-fold compared with that of the ${it E. coli}$ by atomic absorption spectrometer analysis. This result suggested that ${it D. radiodurans}$ and ${it D. grandis}$ are best two suitable species for the development of high Cs-accumulating bacteria by ion beam breeding technology.

論文

Molecular analysis of polyphosphate biosynthesis-related genes in ${it Deinococcus radiodurans}$

上田 涼史郎; 佐藤 勝也; 林 秀謙*; 鳴海 一成*; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 118, 2015/03

It is suggesting a possibility that the polyphosphate granules in cells could accumulate harmful metal ions such as radiocesium to protect the cells from cytotoxicity. ${it Deinococcus radiodurans}$ has been considered as a microorganism for bioremediation under highly radioactive contaminated environments. We generated the polyphosphate biosynthesis-related genes (${it ppk}$ and ${it ppx}$) disruptant strains and characterized their disruption effect to clarify the role of polyphosphate for the accumulation of cesium in ${it D. radiodurans}$. The intracellular levels of polyphosphates in the ${it ppk}$ disruptant and overexpressing strains were almost same and slightly increased, respectively. On other hand, the intracellular level of polyphosphates significantly increased in the ${it ppx}$ disruptant strains, suggesting that the ${it ppx}$gene plays an important role in the accumulation of polyphosphate in ${it D. radiodurans}$.

論文

Genome analysis of the high temperature tolerant mutant of ${it Bradyrhizobium japonicum}$ USDA110 generated by ion-beam irradiation

武田 喜代子*; 佐藤 勝也; 鳴海 一成*; 大野 豊; 大津 直子*; 横山 正*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 120, 2015/03

Major constraint of biofertilizer utilization in agricultural practice is a viability loss of beneficial microorganisms in biofertilizers by high temperature stress during both storage and transportation. Therefore, we have tried to improve high temperature tolerance of ${it Bradyrhizobium japonicum}$ USDA110 using carbon-ion beams accelerated by an AVF cyclotron at TIARA. Consequently, we have obtained a high temperature tolerant mutant that can maintain a high survival rate at 42 $$^{circ}$$C for at least 7 days in yeast-mannitol broth medium, and named it as M14. The genome sequence of M14 was determined by a whole-genome shotgun strategy. The acquirement of high temperature tolerance in M14 could be attributed to 18 base mutations and/or a large-scale inversion (1.27 Mb). In the future, we will conduct further analysis on these mutations in order to decipher the molecular mechanisms of high temperature tolerance of M14.

論文

Thermotolerant mutants of entomopathogenic fungi obtained by ion beam- and $$gamma$$ ray-induced mutagenesis

西東 力*; Fitriana, Y.*; 佐藤 勝也; 大野 豊; 鳴海 一成*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 121, 2015/03

Entomopathogenic fungi, such as ${it Metarhizium anisopliae}$, are economically important agents for integrated pest management programmes of insect pests. However, negative effects of temperature, such as heat stress that exceeds 35 $$^{circ}$$C, can have serious deleterious effects on conidial germination and persistence, vegetative growth, sporulation, and/or the infection process in these entomopathogenic fungi. In this study, we attempted to enhance thermotolerance in a model entomopathogenic fungus, ${it M. anisopliae}$, by mutagenesis using ion beams or $$gamma$$ rays and evaluated the relative virulence of the resulting mutants compared with the wild-type isolates. In conclusion, ion beams and $$gamma$$ rays are useful tools for improving biological characteristics, such as thermotolerance and fungicide-tolerance, in entomopathogenic fungi, but the mutants obtained in this study must be carefully evaluated for unpredictable negative side effects.

論文

Screening of yeast strain for ethanol fermentation after carbon ion beam irradiation

荻野 千秋*; 山田 良祐*; 佐藤 勝也; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 122, 2015/03

Ethanol fermentation from hydrolyzed lignocellulosic biomass with yeast is very important. However, in common sense, there are several chemical inhibitors for yeast from hydrolyzed biomass. Therefore, screening of yeast strain, possessing the tolerance properties against these chemical inhibitors, should be found from nature resources or mutant libraries. In this research, by carbon ion beam irradiation, the screening of candidate yeast strains was performed. Compared with wild type strain, the ethanol production yield was improved in the case of selected strain. It was assumed that the carbon ion beam irradiation could be influence to yeast cell, and improve the metabolic pathway. In future, by using this selected yeast, the ethanol fermentation from hydrolyzed biomass with high efficiency would be performed.

論文

Genome analysis of high ethyl caproate producing sake yeasts generated by ion beam breeding

増渕 隆*; 日向 弘和*; 上田 涼史郎; 林 秀謙*; 池永 裕*; 佐藤 勝也; 大野 豊

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 123, 2015/03

We analyzed genome sequence of the high ethyl caproate producing sake yeast mutant (No.227) generated by ion beam breeding in order to investigated factors contributing the reduced ability to alcohol fermentation. In the high ethyl caproate producing sake yeast, four pyruvate decarboxylase genes (PDC), six alcohol dehydrogenase genes (ADH), two biotin synthesis genes (BIO) and chitinase gene (CTS1) might be involved in the reduced ability to alcohol fermentation. The genome sequence of the strain No.227 was determined by a whole-genome shotgun strategy using pyrosequencing method and compared with the whole-genome sequence of the sake yeast strain Kyokai 7, which is characterized by the fermentation property, as a reference sequence. For the PDC, ADH, BIO and CTS1 loci, no mutation was found in the strain No.227, suggesting that these genes did not involved in the reduced ability to alcohol fermentation.

論文

Role of DNA repair and effect of herbal extract on LOH induced by ion beam radiations in ${it Saccharomyces cerevisiae}$

布柴 達男*; 山内 理子*; 岩田 梨佳*; 佐藤 勝也; 大野 豊; 鳴海 一成*

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 124, 2015/03

The LOH (loss of heterozygosity) induction by various ion beam radiations was investigated in diploid ${it Saccharomyces cerevisiae}$. The ion beams $$^{12}$$C$$^{5+}$$, $$^{12}$$C$$^{6+}$$, $$^{4}$$He$$^{2+}$$ and $$^{40}$$Ar$$^{13+}$$ at the lowest dose of 37.5 Gy, which had $$sim$$10% lethality, induced LOH with $$sim$$5$-fold higher frequency, with dose-dependent manner until 150 Gy. The LOH induction was predominantly ($sim$$90%) resulted from homologous recombination. Pol$$zeta$$ may be involved in the induction of LOH by ion beam radiations, because in the strain lacking Rev3, only slight induction of LOH was observed, whereas no remarkable effect of deletion for Pol$$zeta$$ was observed on ion beam induced LOH. In addition, the effects of ${it Eucalyptus regnans}$ extracts were examined on iron-beam radiation induced LOH, and ethanol extract of ${it Eucalyptus regnans}$ possessed inhibitory effect on LOH induced by $$^{12}$$C$$^{6+}$$, $$^{4}$$He$$^{2+}$$ and $$^{40}$$Ar$$^{13+}$$ion-beam radiation.

論文

Effect of $$gamma$$ radiolysis on pit initiation of Zircaloy-2 in artificial seawater

本岡 隆文; 小松 篤史; 塚田 隆; 山本 正弘

JAEA-Review 2014-050, JAEA Takasaki Annual Report 2013, P. 18, 2015/03

人工海水でのジルカロイ2の孔食発生に対する$$gamma$$線照射による水質変化の影響を調査した。人工海水への$$gamma$$線照射により生成した過酸化水素が不動態被膜を形成し、その不動態被膜により孔食発生が困難になっていることを、溶液分析、電気化学試験並びに表面分析の結果から明らかにした。

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