A quick isolation method for mutants with high lipid yield in oleaginous yeast

A novel method has been developed to easily isolate the mutants with high lipid yield after irradiating oleaginous yeast cells with carbon ions of energy of 80 MeV/u. Pre-selection of the mutants after ion irradiation was performed with culture medium in which the concentration of cerulenin, a potent inhibitor of fatty acid synthetase, was at 8.96 mu mol/l. Afterwards, lipid concentration in the fermentation broth of the pre-selected colonies was estimated by the sulfo-phospho-vanillin reaction instead of the conventional methanol-chloroform extraction. Two mutants with high lipid yield have been successfully selected out by the combined method. This easy and simple method is much less time-consuming but very efficient in the mutant isolation, and it has demonstrated great potential on mutation breeding in oleaginous microorganism.

Induction of cytogenetic adaptive response in spermatogonia and spermatocytes by pre-exposure of mouse testis to low-dose C-12(6+) ions

To investigate the effects of pre-exposure of mouse testis to low-dose C-12(6+) ions on cytogenetics of spermatogonia and spermatocytes induced by subsequent high-dose irradiation. the testes of outbred Kun-Ming strain mice were irradiated with 0.05 Gy of C-12(6+) ions as the pre-exposure dose, and then irradiated with 2 Gy as challenging dose at 4 h after per-exposure. Poly(ADP-ribose) polymerase (PARPs) activity and PARP-1 protein expression were respectively measured by using the enzymatic and Western blot assays at 4 h after irradiation; chromosomal aberrations in spermatogonia and spermatocytes were analyzed by the air-drying method at 8 h after irradiation. The results showed that there was a significant increase in the frequency of chromosomal aberrations and significant reductions of PARP activity and PARP-1 expression level in the mouse testes irradiated with 2 Gy of C-12(6+) ions. However, pre-exposure of mouse testes to a low dose of C-12(6+) ions significantly increased PARPs activity and PARP-1 expression and alleviated the harmful effects induced by a subsequent high-dose irradiation. PARP activity inhibitor 3-aminobenzamide (3-AB) treatment blocked the effects of PARP-1 on cytogenetic adaptive response induced by low-dose C-12(6+) ion irradiation. The data suggest that pre-exposure of testes to a low dose of heavy ions can induce cytogenetic adaptive response to subsequent high-dose irradiation. The increase of PARP-1 protein induced by the low-dose ionizing irradiation may be involved in the mechanism of these observations. (C) 2008 Elsevier B.V. All rights reserved.

Assessment of DNA damage of Lewis lung carcinoma cells irradiated by carbon ions and X-rays using alkaline comet assay

DNA damage and cell reproductive death determined by alkaline comet and clonogenic survival assays were examined in Lewis lung carcinoma cells after exposure to 89.63 MeV/u carbon ion and 6 MV X-ray irradiations, respectively. Based on the survival data, Lewis lung carcinoma cells were verified to be more radiosensitive to the carbon ion beam than to the X-ray irradiation. The relative biological effectiveness (RBE) value, which was up to 1.77 at 10% survival level, showed that the DNA damage induced by the high-LET carbon ion beam was more remarkable than that induced by the low-LET X-ray irradiation. The dose response curves of '' Tail DNA (%)'' (TD) and "Olive tail moment" (OTM) for the carbon ion irradiation showed saturation beyond about 8 Gy. This behavior was not found in the X-ray curves. Additionally, the carbon ion beam produced a lower survival fraction at 2 Gy (SF2) value and a higher initial Olive tail moment 2 Gy (OTM2) than those for the X-ray irradiation. These results suggest that carbon ion beams having high-LET values produced more severe cell reproductive death and DNA damage in Lewis lung carcinoma cells in comparison with X-rays and comet assay might be an effective predictive test even combining with clonogenic assay to assess cellular radio sensitivity

Studies on advantages of heavy ions in radiotherapy compared with gamma-rays

Hepatoma and melanoma cells were exposed to C-12(6+) beams generated by HIRFL facility and gamma-rays and the cell response was studied by colony assays as well as the analysis of RBE of carbon ions was evolved. The survival curves of cells irradiated by heavy ions were different from those of cells irradiated by gamma-rays. And two kinds of cell showed the obvious discrepancy in response to the photon and ion irradiation. The results showed that heavy ions have special physical properties and mighty potency to kill cell in both single and fractional irradiation meanwhile it can kill tumor cells with high radioresistance more efficiently. When involved in clinical therapy, heavy ions will enhance the therapy efficiency and decrease the suffering of patients because it can impair the repair for sublethal damage of cells which can lead to fewer irradiation fractions.

Damage accumulation in gallium nitride irradiated with various energetic heavy ions

In this work a study of damage production in gallium nitride via elastic collision process (nuclear energy deposition) and inelastic collision process (electronic energy deposition) using various heavy ions is presented. Ordinary low-energy heavy ions (Fe+ and Mo+ ions of 110 keV), swift heavy ions (Pb-208(27+) ions of 1.1 MeV/u) and slow highly-charged heavy ions (Xen+ ions of 180 keV) were employed in the irradiation. Damage accumulation in the GaN crystal films as a function of ion fluence and temperature was studied with RBS-channeling technique, Raman scattering technique, scanning electron microscopy (SEM) and transmission electron microscopy (TEM). For ordinary low-energy heavy ion irradiation, the temperature dependence of damage production is moderate up to about 413 K resulting in amorphization of the damaged layer. Enhanced dynamic annealing of defects dominates at higher temperatures. Correlation of amorphization with material decomposition and nitrogen bubble formation was found. In the irradiation of swift heavy ions, rapid damage accumulation and efficient erosion of the irradiated layer occur at a rather low value of electronic energy deposition (about 1.3 keV/nm(3)),. which also varies with irradiation temperature. In the irradiation of slow highly-charged heavy ions (SHCI), enhanced amorphization and surface erosion due to potential energy deposition of SHCI was found. It is indicated that damage production in GaN is remarkably more sensitive to electronic energy loss via excitation and ionization than to nuclear energy loss via elastic collisions.

Defect production in silicon carbide irradiated with Ne and Xe ions with energy of 2.3 MeV/amu

In the present work specimens of mono-crystalline silicon carbide (4H polytype) were irradiated to three successively increasing ion fluences ranging from 7.2 x 10(14) to 6.0 x 10(16) ions/cm(2) (corresponding to the peak displacement damage of 1, 4 and 13 dpa) with Ne and Xe ions respectively with the energy of 2.3 MeV/amu. The irradiated specimens were subsequently annealed at temperatures of 1173 and 1273 K. Defect structure was investigated with transmission electron microscopy (TEM) using a cross-sectional specimen preparation technique. The typical microstructures of the annealed specimens irradiated with Ne or Xe ions to high fluences are characterized by small gas bubbles in high concentration in the peak damage region and black dots and dislocation loops (located in the basal plane) in a shallower and broader depth region. Larger dislocation loops were observed in the Xe-ion irradiated specimen than in the Ne-ion irradiated specimen at the same peak damage level. The enhanced formation of dislocation loops in the case of Xe-ion irradiation is understandable by assuming stronger inclination of heavier inert-gas atoms to occupy substitute site in the peak damage region.

Color center formation in alpha-Al2O3 induced by high energy heavy ions

Single crystals of alpha-alumina were irradiated at room temperature with 1.157 (GeVFe)-Fe-56, 1.755 (GeVXe)-Xe-136 and 2.636 (GeVU)-U-238 ions to fluences range from 8.7 x 10(9) to 6 x 10(12) ions/cm(2). Virgin and irradiated samples were investigated by ultraviolet visible absorption measurements. The investigation reveals the presence of various color centers (F, F+, F-2(2+), F-2(+) and F-2 centers) appearing in the irradiated samples. It is found that the ratio of peak absorbance of F-2 to F centers increases with the increase of the atomic numbers of the incident ions from Fe, Xe to U ions, so do the absorbance ratio of F-2(2+) to F+ centers and of large defect cluster to F centers, indicating that larger defect clusters are preferred to be produced under heavier ion irradiation. Largest color center production cross-section was found for the U ion irradiation. The number density of single anion vacancy scales better with the energy deposition through processes of nuclear stopping, indicating that the nuclear energy loss processes determines the production of F-type defects in heavy ion irradiated alpha-alumina.

Melatonin modulates acute testicular damage induced by carbon ions in mice

The present study was performed to obtain evidence of the radioprotective function of melatonin at different administration levels on carbon ion-induced mouse testicular damage. Outbred Kun-Ming strain mice were divided into six groups, each composed of eight animals: control group, melatonin alone group, irradiation group and three melatonin plus irradiation-treated groups. An acute study was carried out to determine alterations in DNA-single strand break, cell apoptosis, and oxidative stress parameters as well as histopathology in mouse testis 24 h after whole-body irradiation with a single dose of 4 Gy Tie results showed that pre-treatment and post-treatment with high-dose melatonin (10 mg/kg) both significantly alleviated carbon ion-induced acute testicular damage, a greater radioprotective effect being observed in the pre-treatment group. On the other hand, low-dose melatonin (1 mg/kg) had a limited radioprotective effect on irradiation-induced degeneration and DNA lesions in mouse testis. Taken together, the data suggest that prophylactic treatment with a higher dose of melatonin is probably advisable to protect against the effects of heavy-ion irradiation.

Effect of p53 on lung carcinoma cells irradiated by carbon ions or X-rays

The study is to investigate the feasibility and advantages of heavy ion beams on radiotherapy. The cellular cycle and apoptosis, cell reproductive death and p53 expression evaluated with flow cytometry, clonogenic survival assays and Western blot analysis were examined in lung carcinoma cells after exposure to 89.63 MeV/u carbon ion and 6 MV X-ray irradiations, respectively. The results showed that the number colonyforming assay of A549 was higher than that of H1299 cells in two radiation groups; A549 cellular cycle was arrested in G(2)/M in 12 It and the percentage of apoptosis ascended at each time point of carbon ion radiation with doses, the expression of p53 upregulated with doses exposed to X-ray or carbon ion. The cell number in G(2)/M of H1299 and apoptosis were increasing at all time points with doses in C-12(6+) ion irradiation group. The results suggested that the effects of carbon ions or X rays irradiation on lung carcinoma cells were different, C-12(6+) ion irradiation could have more effect on upregulating the expression of p53 than X-ray, and the upregulated expression of p53 might produce the cellular cycle G(2)/M arrested, apoptosis increasing; and p53 gene might affect the lung cancer cells radiosensitivity.

Inactive and mutagenic effects induced by carbon beams of different LET values in a red yeast strain

To evaluate biological action of microorganism exposed to charged particles during the long distance space exploration. Induction of inactivation and mutation in a red yeast strain Rhodotorula glutinis AY 91015 by carbon beams of different LET values (14.9-120 0 keV mu m(-1)) was investigated It was found that survival curves were exponential, and mutation curves were linear for all LET values The dependence of inactivation cross section on LET approached saturation near 120 0 keV mu m(-1) The imitation cross section saturated when LET was higher than 582 keV mu m(-1) Meanwhile, the highest RBEI for inactivation located at 120 0 key mu m(-1) and the highest RBEm for mutation was at 58.2 key mu m(-1) The experiments imply that the most efficient mutagenic part of the depth dose profile of carbon ion is at the plateau region with intermediate LET value in which energy deposited is high enough to Induce mutagenic lesions but too low to induce over kill effect in the yeast cells (C) 2010 Elsevier B V All rights reserved

一氧化氮介导的神经胶质瘤细胞对重离子辐射抗性及动物实验降能装置设计

目的：１、研究一氧化氮(NO)介导的神经胶质瘤细胞对重离子辐射抗性；２、研究小鼠大脑受重离子辐照后的损伤修复；３、为了更好的研究小鼠大脑受重离子布拉格（Bragg）峰区辐照后的损伤修复，设计并制造了旋转轮状降能装置。材料与方法：１、采用兰州重离子研究装置（HIRFL）加速的碳离子束辐照人类神经胶质瘤三种基因型细胞株：野生型神经胶质瘤细胞（A172），带绿色荧光蛋白基因的神经胶质瘤细胞（EA172）和带诱导型一氧化氮合酶基因（iNOS）的神经胶质瘤细胞（iA172），以及一种加了一氧化氮合酶抑制剂（L-NAME）的A172细胞（L-NAME-A172）。分别利用化学法、流式细胞术和MTT法检测三种神经胶质瘤细胞中NO、谷胱甘肽（GSH）的含量，以及它们的细胞周期变化和辐照后存活状况。２、利用不同剂量的碳离子束辐照昆明小鼠全脑，采用八臂迷宫检测随时间推移及训练次数的增加小鼠记忆损伤恢复情况。３、采用蒙特卡罗法和模拟退火法设计制造了一个有机玻璃材料的旋转降能装置。结果：1、NO和GSH在iA172细胞中的含量比A172和EA172中显著要高；辐照后24小时，观察到A172和EA172细胞发生周期阻滞即细胞阻滞于G2／M期，而这种现象没有在iA172细胞中观察到，并且iA172在接受同样辐射刺激后，细胞存活率显著高于其它三个细胞株。2、动物实验表明，在0－2Gy的碳离子辐照在短期内影响小鼠的记忆，经过一定时间后，这种影响得到恢复。3、物理实验显示，降能装置的实验数据与理论计算相符。结论：在低剂量的重离子坪区辐射条件下，一定浓度的NO可以使神经胶质瘤细胞产生辐射抗性。低剂量的重离子辐射致脑损伤可以得到很快的修复

离子辐照生防菌BJ1突变菌株选育及其诱导抗性机制的研究

目的： 利用重离子辐照技术选育出对农作物具有更好防病促生作用的突变生防菌株，探讨利用突变株诱导黄瓜对枯萎病菌产生抗病性的作用机制。 材料与方法： 采用兰州重离子研究装置(HIRFL)加速的碳离子束辐照生防菌BJ1，测定抑菌能力、抑菌谱，确定对该菌株最适宜的离子辐照参数，选育突变菌株。对突变株进行室内盆栽和田间防病促生试验。对原菌株和突变株进行16SrDNA和生理生化反应鉴定，确定分类地位。以突变株为对象进行诱导黄瓜对枯萎病菌产生抗病性的实验。 结果与结论： 1.重离子辐照生防菌BJ1的存活曲线随剂量的增加，呈先降后升再降的马鞍型变化，但是由于离子的能量不同也存在差异，表现为在相同的剂量下，能量越低其能量沉积效应即传能线密度（LET）越大，致死率越高。诱变效果随LET的不同也不尽相同，高LET时的突变株不但有更广的抑菌谱而且抑菌活性较对照也有比较大的提高，在存活率较高的条件下，低剂量就可以得到较多的突变体，有利于筛选优良的正突变体； 2.对于生防菌BJ1最适宜的12C6+辐照参数应选择剂量在200-400Gy，LET为60keV/m范围可筛选获得抑菌活性较高的菌株； 3.通过12C辐照结合抑菌试验最终获得了突变菌株154，该突变株通过20代的移植能够稳定遗传； 4.利用突变株154对黄瓜枯萎病菌进行室内盆栽促生试验，结果表明突变株154能够使促进黄瓜幼苗的生长发育，同时提高了黄瓜植株的抗病性，在对黄瓜枯萎病的防治效果上经154处理的达到了70.34%，高于原菌株和农药防治的效果； 5.传统的生理生化特征结合16SrDNA同源性比对的方法，对菌株BJ1及其突变株154的鉴定结果表明二者均属枯草芽孢杆菌，亲缘关系近，但是突变株154生化测定不同于原菌株，表现为抗菌物质的产量较高； 6.BJ1、154都可在番茄、当归和黄芪的根部有效的定殖，适应根部的生长环境， 并且154的定殖能力稍强； 7.BJ1、154防治当归麻口病效果较好，防治效果最好的是154的20倍液浸苗，防效为82.6%；BJ1的20倍液浸苗与154的10倍液浸苗对麻口病防治效果差别不大，防效分别为78.3%、75.62%；其余处理防效低于62%。 8.BJ1和154处理黄瓜幼苗后，植物体内一系列与抗病性有关的保护酶的活性均有不同程度的提高，因而可认为这些酶活性的改变与生防菌诱导的黄瓜对枯萎病的抗性可能有一定的相关性

经碳离子束诱变的甜高粱汁酒精发酵高产菌株的研究

为了选育出适合发酵甜高粱汁来生产酒精的酵母菌株，本论文以酒精酵母Saccharomyces cerevisiae YY为材料，利用兰州近代物理研究所重离子研究装置（HIRFL）产生的100MeV/u碳离子束对酒精酵母进行了辐照诱变。采用红四氮唑（TTC）作为筛选指示剂，初筛得到了5株产酒能力有所提高的突变酵母菌。通过甜高粱汁发酵，测定发酵液中酒精含量和残糖，复筛出产酒精能力比出发菌株有明显提高的诱变菌株T4。并对其发酵条件进行了优化，以期获得的结果能够为甜高粱汁工业化生产酒精提供参考数据。通过本论文的研究，得到以下初步结果： 1. 在甜高粱汁培养基中，酒精酵母YY的对数生长期在8-20h之间，此时菌体的生长繁殖比较旺盛，活力最佳，为辐照诱变的最佳时期。辐照后，菌体的存活率随辐照剂量的增加呈现出逐渐衰减的趋势。 2. 红四氮唑TTC是一种无色显色指示剂，活菌中所含的脱氢酶可将它还原成红色，因此可以根据菌落呈色的深浅判断酵母菌产酒精能力的高低，从而挑选出产酒能力较高的菌株。本试验用TTC双层培养基法初步筛选出了利用甜高粱汁发酵生产酒精能力较强的T4酵母菌株。 3. 对影响T4菌发酵甜高粱汁生产酒精的几个主要因素（甜高粱汁糖度、接种量、温度、pH、无机盐）进行了初步探讨研究，得出了T4菌发酵甜高粱汁生产酒精的最适条件为：甜高粱汁糖度22%，接种量10%，温度30oC,pH 4.5 ,无机盐加入量为:（NH4）2SO4 1g/L，KH2PO4 5g/L，MgSO4 3g/L。 4. 对发酵条件进行优化后的中试结果显示：出发菌株YY发酵甜高粱汁的时间为36h，酒精产量为8.6% (V/V) ，而T4突变菌甜高粱汁发酵液中的最终酒精含量可以达到9.8%，发酵时间仅为24h。因此，T4菌在工业应用中很有前景

薄膜硅材料的重离子辐照改性研究

选择具有重要应用价值的纳米晶硅及非晶硅薄膜材料，对比研究了快重离子辐照薄膜Si材料所引起的辐照效应，主要有如下工作： 1) 用等离子增强化学气相沉积法(PECVD)以及常压化学气相沉积法(APCVD)在玻璃衬底上制备纳米晶硅薄膜(nc-Si)与非晶硅(a-Si)薄膜。制备的nc-Si和a-Si薄膜的厚度分别为∼200 nm和∼400 nm。 2) 室温下，用94 MeV的Xe离子辐照nc-Si和a-Si薄膜以及单晶硅(c-Si，晶向(100)，电阻率0.001 Ω·cm，作为对比样品)，辐照量分别为1.0×1011、1.0×1012 和1.0×1013 ions/cm2。所有样品均在室温下用UV/VIS/NIR光谱仪、Raman谱仪进行检测分析，对比研究纳米晶、非晶、单晶硅样品的光学性能及结构的变化。 3) 通过UV/VIS/NIR谱，我们得到了不同结构硅材料的光学带隙值及折射率随辐照量的变化规律。通过分析硅材料辐照前后Raman谱的变化，讨论了不同晶体结构的影响。 4) 结合UV/VIS/NIR与Raman结果初步讨论了硅材料结构影响辐照效应的机理。对于a-Si薄膜样品，辐照可能引起了样品局域的再结晶。对于nc-Si薄膜样品，Xe离子辐照先引起nc-Si晶粒的破碎，使样品出现非晶化过程，然后再遵从a-Si薄膜样品中离子辐照效应变化规律

碳离子辐照后细胞的适应性反应及单一时相细胞的相对生物学效应

目的：探讨低剂量碳离子辐照细胞引起的适应性反应,将低剂量效应的研究范围拓宽至高LET电离辐射领域;研究单一时相细胞经低能碳离子辐照后,存活曲线、失活截面和相对生物学效应与细胞周期各时期的关系.材料与方法：采有中国仓鼠肺V79细胞和小鼠黑色素瘤B16细胞,利用兰州近代物理研究所重离子研究装置（HIRFL）产生的碳离子,主要以细胞存活分数和细胞微核率为生物终点,探讨高LET的碳离子对V79细胞和B16细胞的低剂量效应.采用中国仓鼠卵巢细胞CHO-K和细胞同步化方法,利用德国GSI的直线加速器（Unilac）终端产生的低能碳离子,以细胞存活分数为生物终点,研究单一时相细胞经低能碳离子辐照后,存活曲线、失活截面与细胞周期各时期的关系,并与X射线的结果作了比较,得出了各时期细胞的相对生物学效应.结果：低剂量碳离子辐照细胞引起的生物学效应：1.和γ射线一样,低剂量重离子,如0.02Gy的碳离子辐照也能提高细胞的克隆形成率.这种集落形成能力的提高可能与低剂量辐照使细胞对受损DNA的修复能力增强有关.2.从细胞存活分类来看,0.02Gy预处理能使V79和B16两种细胞产生较明显的适应性反应;0.05Gy预处理引起V79细胞的适应性反应不明显,未能引起B16细胞的适应性反应.3.从细胞微核率来看,0.02Gy预处理能引起两种细胞的适应性反应;0.05Gy预处理未能引起两种细胞的适应性反应,并且已表现出协同损伤的趋势.4.高LET的碳离子和低LET射线一样,可以诱导细胞的兴奋效应或适应性反应.但诱导剂量D1不宜过高.