Name: removal of trace elements by cupric oxide nanoparticles from uranium in situ recovery bleed water and its effect on cell viability
Uploaded: 2015-06-21T14:00:00.000+00:00
Duration: 9 min 23 s
Description: Watch this Scientific Journal Video about Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability at JoVE.com

We thank Dr. Roger Hopper and the Wyoming Department of Agriculture, Analytical Services Lab for the mass spectroscopy analysis of our samples. We would like to express our gratitude to the University of Wyoming, School of Pharmacy for allowing us to video this protocol in their laboratories. We would also like to thank the Theodore O. and Dorothy S. King Endowed Professorship Agreement for their graduate assistantship (SC), the University of Wyoming for the Graduate Assistantship support (JRS), and the Science Posse (NSF GK-12 Project # 084129) for the teaching fellowship (JRS). We would also like to thank Uranium One for allowing us to obtain samples and assisting us with questions. This work was supported by the School of Energy Resources, University of Wyoming.

所有样本收集在怀俄明州铀ISR设施铀液体处理建设。 1.生产渗水（PBW） <ol><li>收集两种类型的水样从ISR铀设施:PBW和反渗透（RO）水。从离子交换处理后的监测抽头但反渗透净化之前收集PBW。收集的RO样品后PBW通过反渗透处理去污。 注:浸滤剂输送来自多个井场的铀处理液体的建筑物，在那里被收集在柱和离子交换制得的管道。大约经过离子交换的浸滤剂的1-3％被从电路移除，称为生产排出水（PBW）。 PBW被重新用在挖掘过程或净化/脱钙用RO过滤。 </li><li>收集在高密度聚乙烯水样（HDPE）瓶，根据零头部空间为样品采集和环境质量的怀俄明部门（WYDEQ）23分析标准作业程序。 </li><li>测量温度和pH现场及运输样品在冰上，让他们冷静。 </li><li>商店PBW在4℃。保持PBW液冷却，直至浓缩鹰的最低基本培养基（EMEM-10X），此前有媒体在制备过程中加入的指示在以下协议。 注:PBW是氧化的溶液，将沉淀如果允许冻结或温热至室温。稀释后的PBW溶液足够稀该应用之前它不会沉淀当加热到37℃至细胞和孵育期间。 </li></ol> 2.制备氧化铜纳米粒子（的CuO-NPS） <ol><li>结合含有250mM毫升的0.2M的CuCl 2•2H 2 O，加入250毫升的0.4M的氢氧化钠（NaOH），和5克聚乙二醇纯乙醇溶液（PEG）在一个圆底烧瓶中6毫米硼硅酸盐玻璃球。 </li><li>放置在修改后的微波炉的溶液，并允许它在20％的功率（的6秒的时间间隔，24秒关）在回流下反应，在环境空气压力10分钟。 </li><li>冷却该溶液到室温（20℃），然后倾入50ml锥形管中，留下的玻璃球。 </li><li>离心机在1000×g离心30分钟，滗析该溶液在50ml锥形管中，然后洗的CuO纳米粒用300毫升热水（60-65℃），100毫升乙醇中的序列，和100毫升丙酮。 </li><li>干燥的CuO纳米粒在50ml锥形管中室温（20℃）。 </li><li>刮氧化铜纳米粒他们的管研钵中。覆盖的CuO纳米粒用锡箔和加热的CuO纳米粒至110℃的炉中以除去残留的液体。结合氧化铜纳米粒为一个批次，重量CuO的纳米粒。 注:氧化铜纳米粒和CuO-NP处理PBW的制备是在水中进行的Qual生态系统科学与管理，怀俄明大学的实验室性。的CuO-NP合成接着马丁森和雷迪（2009）11的方法。 </li></ol> 3.治疗PBW与氧化铜纳米粒<ol><li>加50毫克的CuO-NP（1毫克/毫升），以50毫升锥形管中随后50毫升PBW的。密封管，并在250转反应30分钟的长凳上顶轨道摇床。 </li><li>离心样品管在250×g离心30分钟，然后用0.45微米的注射器过滤器过滤上清液。改变离心机速度和时间可以取决于纳米颗粒，以确保的CuO纳米粒成为紧凑的离心管中。 </li></ol> 4.元素分析<ol><li>制备未处理（对照）和CuO-NP处理PBW样品进行元素分析如下。 </li><li>酸化的CuO-NP-处理和未处理PBW等份（40毫升）与痕量金属级硝酸至pH 2.0。对于阳离子通过感应coupl分析酸化PBW等分编等离子体-质谱（ICP-MS），如雷迪和Roth（2012）13中所述。 </li><li>制备的CuO-NP-处理和未处理PBW的未酸化等份（20毫升）并分析未酸化等份阴离子通过如雷迪和Roth（2012）13中所述的离子色谱法（IC）。 注:等分被农业部分析服务的怀俄明部门，怀俄明州拉勒米的82070. IC和等离子体质谱过程的描述可以在雷迪和罗斯，（2012）13中找到分析。 </li></ol> 5.制备细胞培养基使用PBW <ol><li>使用两个控制（EMEM-1x和RO +媒体）和八个PBW测试的媒体解决方案（四种浓度每未处理PBW和CuO-NP处理的介质）的可行性研究。是溶液的概述如下: <ol><li>对于EMEM-1X控制，购买的Eagle氏最低必需培养基（EMEM-1×）与L-谷氨酰胺和碳酸氢钠已经添加。添加胎牛血清（FBS）并且按照制造商的说明，抗生素。 注:EMEM-1X被购买稀释到适当的浓度对细胞生长和含L-谷氨酰胺和碳酸氢钠。 EMEM-1X需要添加胎牛血清（FBS）和青霉素和链霉素（50IU / ml青霉素和50μg/ ml链霉素）的抗生素混合物的。 EMEM-1X被用作对照培养基，因为它是在制造商推荐的生长培养基在该研究中使用两种类型的细胞。浓缩EMEM-10倍，从设施或未经处理或的CuO-NP处理PBW稀释RO水，以产生测试溶液。浓缩EMEM-10倍时购买不包含所以这些被添加除了胎牛血清（FBS）和青霉素和链霉素的抗生素混合物L-谷氨酰胺或碳酸氢钠。 </li><li>对于RO控制解决方案中使用RO水从ISR收集设施。使用相同的协议PBW测试仅媒体替代100％RO笏呃从地方PBW的ISR设施。从实验室稀未处理和CuO-NP处理液使用的RO或超纯水。 </li><li>稀未处理PBW为四个测试浓度与细胞培养基成分混合之前。通过在下列组合混合未处理PBW用RO（从实验室）制备的4种不同浓度的未处理PBW溶液:100％（纯PBW +无RO水），75％（187.5毫升PBW +62.5毫升RO水的）， 50％（125毫升PBW + 125毫升RO水的）或25％（62.5毫升PBW +187.5毫升的RO水）。 </li><li>稀的CuO-NP处理PBW为四个测试浓度与细胞培养基成分混合之前。在下列组合通过混合PBW制备的4种不同浓度的的CuO-NP处理PBW溶液（预处理用1毫克/毫升的CuO-NP，30分钟）用RO（从实验室）:100％（纯CuO- NP处理PBW +无RO水），75％（187.5毫升的PBW +62.5毫升RO水的CuO-NP处理的），50％（125毫升的CuO-NP处理PBW + 125毫升RO水）或25％（62.5毫升的CuO-NP处理PBW +187.5毫升的RO水）。 </li></ol></li><li>加入25毫升浓EMEM-10倍的190毫升100％的RO及100％，75％，50％或准备250毫升反渗透+媒体，未处理PBW +媒体和CuO-NP处理PBW +媒浓度在步骤6.1.3和6.1.4中创建的预制未处理的或氧化铜-NP处理PBW浓度的25％。 </li><li>调节每个溶液至7.4，用NaOH或HCl的pH。 </li><li>补充未处理和CuO-NP处理PBW的各个浓度以及反渗透+媒体具有以下标准组件:25毫升（10％）胎牛血清（FBS），2.5毫升L-谷氨酰胺，0.55克碳酸氢钠和1.25ml笔/链霉素（50IU / ml青霉素和50μg/ ml链霉素）。 </li><li>调整未处理PBW +介质的每个浓度的渗透压，加入RO水，并用渗透压计测量的CuO-NP处理PBW +媒体以及RO +媒体290-310毫渗透摩尔浓度/千克。 </li><li>使用过滤器每个解决方案0.22微米真空过滤器单元，并储存在4℃。 注意:由于在RO水用于调节渗透压的量的轻微变化，变化在5％范围内的最终介质浓度，与未处理PBW +媒浓度在56％，44％，29％和16.5％，和CuO-NP-处理PBW +媒浓度为53％，45％，30％及17％。 </li></ol> 6.细胞活力注:由于肾脏和肝脏的重金属毒性的靶器官，采用培养的人类胚胎肾（HEK293）细胞（HEK）和人肝癌（肝癌）细胞（HEP）的测试方法24-26。 <ol><li>电镀在根据制造商的说明，将实验中使用的96孔板前2-3天制备HEK和HEP细胞的培养物。 </li><li>使用3- [4，5-二甲基吡啶-2-基] -2，5-二苯基溴化物（MTT）测定法测量细胞活力。 注:MTT法协议是从眉修改rloo 等人。（2011）27。 <ol><li>以粉末形式获得MTT。添加磷酸盐缓冲液（PBS），以弥补股票浓度为50毫克/毫升。搅拌该溶液2小时，然后过滤用0.45μm针筒式滤器和分装成1.5毫升冷冻安全管。避光并储存管在4℃下。 </li></ol></li><li>从用胰蛋白酶，离心其培养皿除去HEK和Hep细胞在1000×g离心5分钟，并滗出胰蛋白酶。加入5毫升的PBS和混合细胞以获得单细胞溶液。然后，应用将20μl单细胞溶液到血球以获得每毫升溶液细胞计数。再次离心细胞在1,000×g离心5分钟，倒出PBS用于冲洗细胞。添加EMEM-1倍的适量调节细胞浓度至500个细胞/ 100微升（100μl/孔）。 </li><li>填充板的周边的孔用200μlPBS中，以控制蒸发。 </li><li>种子细胞s的500个细胞/孔的密度，加入100微升至每个孔，除了外围孔（其不与细胞铺板）。 注:播种密度和HEK Hep细胞是基于实验的增长曲线，使经济增长的高峰期发生天左右4-5。准备生长曲线对所有细胞系来估计接种密度。 </li><li>孵育细胞24小时，在37℃下使它们进行细胞密度的基线MTT读数之前恢复（形式​​紧粘连至板）。 </li><li>通过从第一塔提取播种介质（不包括周边）并加入100微升的MTT（5mg / ml的在介质）向孔1小时进行细胞密度的基线MTT读数。 </li><li>一小时后，取出MTT法和加入100微升二甲基亚砜（DMSO）的溶解由活细胞（20分钟）产生的MTT-甲。 </li><li>读出的第一列的光密度（OD）在570纳米的吸收波长，得到基行读取。 <ol><li>使用基准读数，以确保所有平板接种正确和细胞板之间持续增长。从塔中取出DMSO培养未来24小时前进行测试。 注意:如果二甲基亚砜留在板过夜它拉水分从相邻列，导致在介质体积的减少。 </li></ol></li><li>温暖的测试解决方案（ 即 EMEM-1X，RO，未经处理的PBW和CuO-NP处理PBW媒体解决方案），以37℃的水浴。 </li><li>从板（不包括周边或将其用于基线读数的第一列），并用100微升EMEM-1倍，反渗透+媒体，未处理PBW +媒浓度或的CuO-NP的取代的其余部分移除播种媒体 - 处理PBW +媒体浓度（每盘一个解决方案）。孵育细胞在他们的测试浓度或控制解决方案，共七天（天2-8）。 注:10共板块:1 EMEM-1X，1 RO +媒体，各1个未处理PBW +媒体浓度（56％，44％，29％和16.5％），每个的CuO-NP处理PBW +媒体集中的一个板块（53％，45％ ，每细胞系实验的30％和17％）。 </li><li>每天以下基准MTT阅读，从各自板的下一列取下控制和测试解决方案（下6.11的说明中列出）（ 如第2天测试和控制媒体从第3行中删除，BG井;第3天:行4，井BG 等 ），并重复MTT协议为在上述步骤6.7-6.9所述。 </li><li>每天重复协议七天。平均外径结果的每一行（6孔），并报告与时间，以产生一个七天的生长曲线。 </li><li>为了评估铜螯合的对细胞活力的影响的CuO-NP处理PBW +媒体按照相同的步骤如上述，除了加入的解决方案，以它们各自的板之前添加的D-青霉胺100μM的控制和测试方案。执行数据肛ysis用科学绘图软件。 </li></ol> 7.地球化学模拟<ol><li>从以下网站下载的Visual MINTEQ 3.0 / 3.1一个免费<a href="http://www2.lwr.kth.se/English/Oursoftware/vminteq/">http://www2.lwr.kth.se/English/Oursoftware/vminteq/</a> 。 注:Visual MINTEQ是一个免费的化学平衡模型的金属形态，溶解平衡，吸附等天然水域的计算。此外，它是用来预测离子物种，离子活动，离子络合物和饱和指数是相对于元件的前，后处理（质谱结果）的浓度，以检查元件除去28的可能机制。 </li><li>打开程序和输入从第4步，包括pH值，碱度和不同的元素的浓度，进入程序的质谱数据。 注:由于地下水的过程中就地 urani氧化嗯提取工艺，用砷，钒和铀的氧化物质为输入。 </li></ol> 8.抑制浓度50（IC 50） <ol><li>由第一平均上的三个独立的运行5天的存活率（OD平均值）计算的IC 50为未处理和CuO-NP处理PBW +介质的浓度。 </li><li>减去第五天从生存能力EMEM-1X的第五天生存能力平均未处理和CuO-NP处理PBW +媒体浓度的平均值来计算可行性的差异。然后除以在EMEM第5天的存活率差异由平均存活率，再乘以100得到的抑制百分比。 </li><li>减去100（EMEM-1X生存能力）抑制百分比，以获取每个未处理和CuO-NP处理PBW +媒体集中的百分比生存能力。 </li><li>输入到通过设置EMEM-1x在一个的浓度和为100％的存活率科学绘图软件;全部改造成浓度日志规模（X =日志（X）），并进行非线性最小二乘回归拟合分析。 </li></ol> 9.数据分析<ol><li>在未经处理的和CuO-NP处理PBW元素的浓度与双尾成对，学生T检验进行比较。 </li><li>通过使用收集的七天增长曲线的数据计算的曲线（AUC）下面积，并分析与重复测量方差分析（ANOVA）的方差，然后所有组之间Tukey的事后比较（n = 3时）。 </li><li>通过使用数据从生长曲线的五天为未处理和CuO-NP处理PBW +媒体溶液（如上所述）计算IC 50值。对&lt;0.05被认为是显著P值。 注:统计分析的目的，一半的检出限质谱值被分配到离子浓度水平低于该限制29。 </li></ol>

<ol>
	<li>Qu, X., Alvarez, P., Li, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Applications+of+nanotechnology+in+water+and+wastewater+treatment.">Applications of nanotechnology in water and wastewater treatment.</a> Water Research. 47 (12), 3931-3946 (2013).</li><li>Martinson, C., Reddy, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adsorption+of+arsenic(III)+and+arsenic(V)+by+cupric+oxide+nanoparticles.">Adsorption of arsenic(III) and arsenic(V) by cupric oxide nanoparticles.</a> Journal of Colloid and Interface Science. 336 (2), 401-411 (2009).</li><li>Reddy, K., McDonald, K., King, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel+arsenic+removal+process+for+water+using+cupric+oxide+nanoparticles.">A novel arsenic removal process for water using cupric oxide nanoparticles.</a> Journal of Colloid and Interface Science. 397, 96-102 (2013).</li><li>Reddy, K., Roth, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+Removal+from+Natural+Groundwater+Using+Cupric+Oxide.">Arsenic Removal from Natural Groundwater Using Cupric Oxide.</a> Ground Water. 51 (1), 83-91 (2012).</li><li>Zhang, G., Ren, Z., Zhang, X., Chen, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanostructured+iron(III)-copper(II)+binary+oxide:+a+novel+adsorbent+for+enhanced+arsenic+removal+from+aqueous+solutions.">Nanostructured iron(III)-copper(II) binary oxide: a novel adsorbent for enhanced arsenic removal from aqueous solutions.</a> Water Research. 47 (12), 4022-4031 (2013).</li><li>Ali, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+generation+adsorbents+for+water+treatment.">New generation adsorbents for water treatment.</a> Chemical Reviews. 112 (10), 5073-5091 (2012).</li><li>Zhang, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CuO+nanostructures:+Synthesis,+characterization,+growth+mechanisms,+fundamental+properties,+and+applications.">CuO nanostructures: Synthesis, characterization, growth mechanisms, fundamental properties, and applications.</a> Progress in Materials Science. 60, 208-337 (2014).</li><li>Schmidt, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=TOX+21:+new+dimensions+of+toxicity+testing.">TOX 21: new dimensions of toxicity testing.</a> Environmental health perspectives. 117 (8), 348-353 (2009).</li><li>Firestone, M., Kavlock, R., Zenick, H., Kramer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+U.S.+Environmental+Protection+Agency+Strategic+Plan+for+Evaluating+the+Toxicity+of+Chemicals.">The U.S. Environmental Protection Agency Strategic Plan for Evaluating the Toxicity of Chemicals.</a> Journal of Toxicology and Environmental Health, Part B. 13 (2-4), 139-162 (2010).</li><li>Bae, D., Gennings, C., Carter, W., Yang, R., Campain, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toxicological+interactions+among+arsenic,+cadmium,+chromium,+and+lead+in+human+keratinocytes.">Toxicological interactions among arsenic, cadmium, chromium, and lead in human keratinocytes.</a> Toxicological Sciences: An Official Journal of the Society of Toxicology. 63 (1), 132-142 (2001).</li><li>Whittaker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exposure+to+Pb,+Cd,+and+As+mixtures+potentiates+the+production+of+oxidative+stress+precursors:+30-day,+90-day,+and+180-day+drinking+water+studies+in+rats.">Exposure to Pb, Cd, and As mixtures potentiates the production of oxidative stress precursors: 30-day, 90-day, and 180-day drinking water studies in rats.</a> Toxicology and Applied Pharmacology. 254 (2), 154-166 (2011).</li><li>Schilz, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Investigating the ability of cupric oxide nanoparticles to adsorb metal contaminants from uranium in-situ recovery (ISR) production bleed water and assessing the associated changes in cytotoxicity. , (2014).</li><li>Florea, A., Splettstoesser, F., B&#252;sselberg, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+trioxide+(As2O3)+induced+calcium+signals+and+cytotoxicity+in+two+human+cell+lines+SY-5Y+neuroblastoma+and+293+embryonic+kidney+(HEK).">Arsenic trioxide (As2O3) induced calcium signals and cytotoxicity in two human cell lines SY-5Y neuroblastoma and 293 embryonic kidney (HEK).</a> Toxicology and Applied Pharmacology. 220 (3), 292-301 (2007).</li><li>Mao, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cadmium+induces+apoptosis+in+human+embryonic+kidney+(HEK)+293+cells+by+caspase-dependent+and+-independent+pathways+acting+on+mitochondria.">Cadmium induces apoptosis in human embryonic kidney (HEK) 293 cells by caspase-dependent and -independent pathways acting on mitochondria.</a> Toxicology in Vitro. 21 (3), 343-354 (2007).</li><li>Tchounwou, P., Yedjou, C., Patlolla, A., Sutton, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Heavy Metal Toxicity and the Environment. Molecular, Clinical and Environmental Toxicology. 101, 133-164 (2012).</li><li>Meerloo, J., Kaspers, G., Cloos, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+Sensitivity+Assays:+The+MTT+Assay.">Cell Sensitivity Assays: The MTT Assay.</a> Cancer Cell Culture. 731, 237-245 (2011).</li><li>Gustafsson, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Visual MINTEQ. , (2010).</li><li>Hallab, N., Caicedo, M., McAllister, K., Skipor, A., Amstutz, H., Jacobs, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Asymptomatic+prospective+and+retrospective+cohorts+with+metal-on-metal+hip+arthroplasty+indicate+acquired+lymphocyte+reactivity+varies+with+metal+ion+levels+on+a+group+basis.">Asymptomatic prospective and retrospective cohorts with metal-on-metal hip arthroplasty indicate acquired lymphocyte reactivity varies with metal ion levels on a group basis.</a> Journal of Orthopaedic Research. 31 (2), 173-182 (2013).</li><li>Goswami, A., Raul, P., Purkait, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+adsorption+using+copper+(II)+oxide+nanoparticles.">Arsenic adsorption using copper (II) oxide nanoparticles.</a> Chemical Engineering Research and Design. 90 (9), 1387-1396 (2011).</li><li>Pillewan, P., Mukherjee, S., Roychowdhury, T., Das, S., Bansiwal, A., Rayalu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Removal+of+As(III)+and+As(V)+from+water+by+copper+oxide+incorporated+mesoporous+alumina.">Removal of As(III) and As(V) from water by copper oxide incorporated mesoporous alumina.</a> Journal of Hazardous Materials. 186 (1), 367-375 (2011).</li><li>Kroll, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cytotoxicity+screening+of+23+engineered+nanomaterials+using+a+test+matrix+of+ten+cell+lines+and+three+different+assays.">Cytotoxicity screening of 23 engineered nanomaterials using a test matrix of ten cell lines and three different assays.</a> Particle and fibre toxicology. 8 (9), 1-19 (2011).</li><li>Fahmy, B., Cormier, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper+oxide+nanoparticles+induce+oxidative+stress+and+cytotoxicity+in+airway+epithelial+cells.">Copper oxide nanoparticles induce oxidative stress and cytotoxicity in airway epithelial cells.</a> Toxicology in vitro: an international journal published in association with BIBRA. 23 (7), 1365-1371 (2009).</li><li>Radike, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distribution+and+accumulation+of+a+mixture+of+arsenic,+cadmium,+chromium,+nickel+and+vanadium+in+mouse+small+intestin,+kidney,+pancreas,+and+femur+following+oral+administration+in+water+or+feed.">Distribution and accumulation of a mixture of arsenic, cadmium, chromium, nickel and vanadium in mouse small intestin, kidney, pancreas, and femur following oral administration in water or feed.</a> Journal of Toxicology and Environmental Health, Part A. 65 (23), 2029-2052 (2002).</li><li>Barbier, O., Jacquillet, G., Tauc, M., Cougnon, M., Poujeol, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+heavy+metals+on,+and+handling+by,+the+kidney.">Effect of heavy metals on, and handling by, the kidney.</a> Nephron. Physiology. 99 (4), 105-110 (2005).</li><li>Zheng, X., Watts, G., Vaught, S., Gandolfi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Low-level+arsenite+induced+gene+expression+in+HEK293+cells.">Low-level arsenite induced gene expression in HEK293 cells.</a> Toxicology. 187 (1), 39-48 (2003).</li><li>Li, Z., Piao, F., Liu, S., Wang, Y., Qu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subchronic+exposure+to+arsenic+trioxide-induced+oxidative+DNA+damage+in+kidney+tissue+of+mice.">Subchronic exposure to arsenic trioxide-induced oxidative DNA damage in kidney tissue of mice.</a> Experimental and Toxicologic Pathology. 62 (5), 543-547 (2010).</li><li>Farombi, E., Akintunde, J., Nzute, N., Adedara, I., Arojojoye, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Municipal+landfill+leachate+induces+hepatotoxicity+and+oxidative+stress+in+rats.">Municipal landfill leachate induces hepatotoxicity and oxidative stress in rats.</a> Toxicology and Industrial Health. 28 (6), 532-541 (2011).</li><li>Das, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Arsenic+exposure+through+drinking+water+increases+the+risk+of+liver+and+cardiovascular+diseases+in+the+population+of+West+Bengal.">Arsenic exposure through drinking water increases the risk of liver and cardiovascular diseases in the population of West Bengal.</a> India. BMC public health. 12 (1), 639-648 (2012).</li><li>Valko, M., Morris, H., Cronin, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Metals,+toxicity+and+oxidative+stress.">Metals, toxicity and oxidative stress.</a> Current Medicinal Chemistry. 12 (10), 1161-1208 (2005).</li><li>Horie, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+Adsorption+of+Ultrafine+Metal+Oxide+and+Its+Influence+on+Cytotoxicity+toward+Cultured+Cells.">Protein Adsorption of Ultrafine Metal Oxide and Its Influence on Cytotoxicity toward Cultured Cells.</a> Chemical Research in Toxicology. 22 (3), 543-553 (2009).</li></ol>

The authors have nothing to disclose.

以前的研究报告说，氧化铜，纳米粒子去除砷地下水11,13,30,31。这项研究支持，这些以往的调查结果也报告说，氧化铜纳米粒删除PBW更多的污染物。本研究还证实了以前的报告的CuO纳米粒是在除砷有效，尽管其他污染物和可能的竞争离子11的存在。形态建模预测，钒物种PBW的97％的负电荷，从而允许吸附在CuO纳米粒和批量处理除去钒的92％。 这是第一次研究，?...

大约是美国电力供应的20％是由核能，部分基于国家鼓励提高能源独立性提供，预计美国的核能力，以增加1。核能的全球增长预计也将继续，与许多美国以外的2出现的增长。截至2013年，美国铀的83％是进口的，但储量952544吨，在美国3,4存在。在2013年有7个新设施的应用和怀俄明州，新墨西哥州，内布拉斯加州和5之间14重启/扩展应用。在美国，铀为主通过原位恢复（ISR）处理6萃取。 ISR导致更少的土地破坏和避免造成尾矿堆，可以释放环境污染物7。 ISR使用水基氧化解决方案，以从​​地下矿石体，在这之后的铀从浸出液通过提取浸出铀离子交换过程8。维持负面水平衡在矿体，浸出液的一部分，被称为生产流血水（PBW）中，放血关闭。所述PBW的一部分被利用反渗透（RO）和净化重新引入开采过程中，但PBW也可以有利于工业或农业用途，如果有毒的污染物可以减少到由状态管理机构对于表面确定的可接受水平和地下水9。目前，大多数ISR铀的设施使用RO从PBW去除污染物。然而，反渗透处理是能源密集型，并产生有毒废物盐水，这需要监管的处置。 许多水净化方法都存在，包括吸附剂，膜和离子交换。在这些中，吸附是最常用的，并且最近的发展纳米颗粒合成具有增强的吸附剂基水净化处理10的功能。铜OXI德纳米颗粒（氧化铜纳米粒）以前没有被广泛研究对铀的ISR PBW，但在最近的污染物去除从地下水的研究，氧化铜纳米粒被发现有独特的性能，包括无需预处理或后处理的水处理工序（ 例如 ，调节pH值或氧化还原电位），并在不同的水成分表现良好（ 例如 ，在不同的pH值，盐的浓度，或竞争离子）11。另外，氧化铜纳米粒容易通过用氢氧化钠（NaOH），之后，将再生的CuO纳米粒可重复使用浸出再生。从天然水域的CuO-NP微量金属过滤功能细节已先前公布11-14。 虽然对水处理是有用的，金属氧化物纳米颗粒可能是有毒的活生物体，但毒性的程度取决于，部分地基于纳米粒子的特性和成分10,15,16。因此，研究SIMULT是很重要现场应用之前aneous污染物的去除和纳米粒子毒性。目前的研究确定的CuO纳米粒的能力以去除PBW优先污染物（包括砷，硒，钒和铀），和评估的CuO-NP处理对PBW细胞毒性的影响。 PBW从有源ISR铀设施收集并用于确定在优先的污染物去除的CuO-NP处理的功效。之前和之后的CuO-NP处理PBW细胞毒性也进行了评估。 PBW是一个复杂的地质（工业/环境）的混合物和环境都和健康科学（NIEHS）国家研究所和机构的有毒物质及疾病登记（ASTDR）是把重点放在学习环境相关的混合物，包括混合物的毒性因为它们在自然界或工业设置，以及在体外测试促进存在优先的化学品进行进一步的体内测试17-19。是慢性，低剂量暴露的混合物具有挑战性的研究，因为长期接触低剂量的混合物，不会产生明显的影响，至少在大多数实验室研究的很短的时间框架。同样，大多数在化学混合物的体外研究细胞暴露于两种或更多种金属20,21限定的实验室制造的混合物。这些研究提供基线信息，但简化的混合物不复制，可能会出现一个天然的，环境样品，其中所述全方位混合物组分都存在于该复合拮抗和协同相互作用。 本研究的目标是检查供PBW备用污染物去除工艺和评估（的CuO-NP）的治疗上PBW细胞毒性使用培养的人类细胞的效果。该结果可能通过更有效的或环保的方法来去除污染物的发展中受益的铀业。这项研究提供第一个证据是PBW减少污染物优先由氧化铜纳米粒在哺乳动物细胞中22毒性降低。

<table><tbody><tr><td>CuCl&lt;sub&gt;2&lt;/sub&gt;</td><td>Sigma</td><td>203149</td><td></td></tr><tr><td>Borosilicate glass balls</td><td>VWR</td><td>26396-639</td><td>6 mm</td></tr><tr><td>Nitric Acid</td><td>Fisher</td><td>A509-P500</td><td>Trace metal grade</td></tr><tr><td>0.45 &amp;#956;m syringe filter</td><td>Fisher</td><td>SLHA 033S S</td><td></td></tr><tr><td>10x EMEM</td><td>Fisher</td><td>BW12-684F</td><td></td></tr><tr><td>Fetal Bovine Serum</td><td>ATCC</td><td>30-2020</td><td></td></tr><tr><td>L-glutamine</td><td>Fisher</td><td>BP379-100</td><td></td></tr><tr><td>NaHCO&lt;sub&gt;3&lt;/sub&gt;</td><td>Sigma</td><td>S5761</td><td></td></tr><tr><td>Penicillin/Streptomycin</td><td>ATCC</td><td>30-2300</td><td></td></tr><tr><td>0.22 &amp;#956;m vacuum filter unit</td><td>Fisher</td><td>09-740-28C</td><td></td></tr><tr><td>HEK293</td><td>ATCC</td><td>CRL-1573</td><td></td></tr><tr><td>HEPG2</td><td>ATCC</td><td>HB-8065</td><td></td></tr><tr><td>Trypsin</td><td>Sigma</td><td>SV3003101</td><td></td></tr><tr><td>MTT</td><td>Sigma</td><td>M2128</td><td></td></tr><tr><td>D-penicillamine</td><td>Fisher</td><td>ICN15180680</td><td></td></tr><tr><td>96-well plates</td><td>Fisher</td><td>07-200-92</td><td></td></tr><tr><td>DMSO</td><td>Fisher</td><td>D12814</td><td></td></tr><tr><td>Spectra Max 190</td><td>Molecular Devices</td><td></td><td></td></tr><tr><td>Visual MINTEQ version 3.0</td><td>KTH Royal Institute of Technology</td><td></td><td></td></tr><tr><td>ICP-MS</td><td>Agilent</td><td></td><td>Details of instruments, models and detection limits were published in Reddy &lt;em&gt;et al.&lt;/em&gt;, 2013.</td></tr><tr><td>IC DIONEX DX 500</td><td>Dionex</td><td></td><td>Details of instruments, models and detection limits were published in Reddy &lt;em&gt;et al.&lt;/em&gt;, 2013.</td></tr><tr><td>VWR Incubator</td><td>VWR</td><td></td><td></td></tr></tbody></table>

removal of trace elements by cupric oxide nanoparticles from uranium in situ recovery bleed water and its effect on cell viability

在原位恢复（ISR）是提取铀在美国的主要方法。期间的ISR，铀是从矿体浸出，并通过离子交换萃取。所得的生产泌水（PBW）含有污染物，如砷等重金属。 PBW从激活ISR铀设施处理样品与氧化铜纳米粒子（氧化铜纳米粒）。的CuO-NP处理PBW降低优先污染物，包括砷，硒，铀，和钒。未处理和CuO-NP处理通过MTT（3-（4,5-二甲基吡啶-2-基）-2,5-二苯基溴化）测定PBW用作细胞生长介质和变化生存能力的液体成分分析在人胚肾（HEK 293）和人肝细胞癌（Hep G2细胞）细胞。的CuO-NP处理用改进的HEK和HEP细胞活力有关。这种方法的限制包括稀释PBW由生长培养基成分和渗透压克分子期间先进而精湛调整以及必要pH调节。这种方法是在其更广泛的范围，由于稀释作用以及变化PBW的pH是传统微酸但是限制;这种方法可以有更广泛的用途评估的CuO-NP处理的较为中性的水域。

PBW成分浓度和pH值在未处理和CuO-NP处理PBW报道于表1中 。马丁森和雷迪（2009）报告说，估计在9.4±0.4的零电荷的CuO-NP的点。鉴于PBW的pH为7.2-7.4，在这些条件下，水捐赠质子的CuO纳米粒，使纳米颗粒表面带正电荷，允许带负电荷的物质的吸附。从PBW的CuO-NP处理去除优先污染物，包括砷，硒，铀和钒（ 表1）。平均砷浓度降低了87％[从0.0175到0.002毫克/升（双尾配对t检验，p</...

Watch this Scientific Journal Video about Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability at JoVE.com

Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

去除微量元素的氧化铜纳米粉体制备铀<em&gt;原位</em&gt;恢复泌水及其对细胞活力的影响

In situ recovery (ISR) is the predominant method of uranium extraction in the United States. During ISR, uranium is leached from an ore body and extracted ...

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Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

9.7K 次观看.  University of New Mexico. Production bleed water (PBW) was treated with cupric oxide nanoparticles (CuO-NPs) and cellular toxicity was assessed in cultured human cells. The goal of this protocol was to integrate the native environmental sample into a cell culture format assessing the changes in toxicity due to CuO-NP treatment.

视频: Removal of Trace Elements by Cupric Oxide Nanoparticles from Uranium In Situ Recovery Bleed Water and Its Effect on Cell Viability

Preparation and Testing of Impedance-based Fluidic Biochips with RTgill-W1 Cells for Rapid Evaluation of Drinking Water Samples for Toxicity

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity sensor. A monolayer of RTgill-W1 cells forms on the sensing electrodes enclosed within the biochips. The biochips are then used for testing in a field portable electric cell-substrate impedance sensing (ECIS) device designed for rapid toxicity testing of drinking water. The manuscript further describes how to run a toxicity test using the prepared biochips. A control water sample and the test water sample are mixed with pre-measured powdered media and injected into separate channels of the biochip. Impedance readings from the sensing electrodes in each of the biochip channels are measured and compared by an automated statistical software program. The screen on the ECIS instrument will indicate either "Contamination Detected" or "No Contamination Detected" within an hour of sample injection. Advantages are ease of use and rapid response to a broad spectrum of inorganic and organic chemicals at concentrations that are relevant to human health concerns, as well as the long-term stability of stored biochips in a ready state for testing. Limitations are the requirement for cold storage of the biochips and limited sensitivity to cholinesterase-inhibiting pesticides. Applications for this toxicity detector are for rapid field-portable testing of drinking water supplies by Army Preventative Medicine personnel or for use at municipal water treatment facilities.

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial cells for use in a field portable electric cell-substrate impedance sensor. The protocol for running a rapid drinking water toxicity test with the sensor is also described.

准备和测试阻抗为基础与RTgill-W1细胞饮用水进行快速评估水样进行毒性射流生物芯片

This manuscript describes how to prepare fluidic biochips with Rainbow trout gill epithelial (RTgill-W1) cells for use in a field portable water toxicity ...

科研

环境科学

Detection and Recovery of Palladium, Gold and Cobalt Metals from the Urban Mine Using Novel Sensors/Adsorbents Designated with Nanoscale Wagon-wheel-shaped Pores

Developing low-cost, efficient processes for recovering and recycling palladium, gold and cobalt metals from urban mine remains a significant challenge in industrialized countries. Here, the development of optical mesosensors/adsorbents (MSAs) for efficient recognition and selective recovery of Pd(II), Au(III), and Co(II) from urban mine was achieved. A simple, general method for preparing MSAs based on using high-order mesoporous monolithic scaffolds was described. Hierarchical cubic Ia3d wagon-wheel-shaped MSAs were fabricated by anchoring chelating agents (colorants) into three-dimensional pores and micrometric particle surfaces of the mesoporous monolithic scaffolds. Findings show, for the first time, evidence of controlled optical recognition of Pd(II), Au(III), and Co(II) ions and a highly selective system for recovery of Pd(II) ions (up to ~95%) in ores and industrial wastes. Furthermore, the controlled assessment processes described herein involve evaluation of intrinsic properties (e.g., visual signal change, long-term stability, adsorption efficiency, extraordinary sensitivity, selectivity, and reusability); thus, expensive, sophisticated instruments are not required. Results show evidence that MSAs will attract worldwide attention as a promising technological means of recovering and recycling palladium, gold and cobalt metals.

Because of the importance and extensive use of palladium, gold and cobalt metals in high-technology equipment, their recovery and recycling constitute an important industrial challenge. The metal recovery system described herein is a simple, low-cost means for the effective detection, removal, and recovery of these metals from the urban mine.

检测和钯的回收，从城市矿山黄金和钴金属使用标有纳米货车车轮形的孔隙新型传感器/吸附剂

Developing low-cost, efficient processes for recovering and recycling palladium, gold and cobalt metals from urban mine remains a significant challenge in ...

工程学

Speciation and Bioavailability Measurements of Environmental Plutonium Using Diffusion in Thin Films

The biological uptake of plutonium (Pu) in aquatic ecosystems is of particular concern since it is an alpha-particle emitter with long half-life which can potentially contribute to the exposure of biota and humans. The diffusive gradients in thin films technique is introduced here for in-situ measurements of Pu bioavailability and speciation. A diffusion cell constructed for laboratory experiments with Pu and the newly developed protocol make it possible to simulate the environmental behavior of Pu in model solutions of various chemical compositions. Adjustment of the oxidation states to Pu(IV) and Pu(V) described in this protocol is essential in order to investigate the complex redox chemistry of plutonium in the environment. The calibration of this technique and the results obtained in the laboratory experiments enable to develop a specific DGT device for in-situ Pu measurements in freshwaters. Accelerator-based mass-spectrometry measurements of Pu accumulated by DGTs in a karst spring allowed determining the bioavailability of Pu in a mineral freshwater environment. Application of this protocol for Pu measurements using DGT devices has a large potential to improve our understanding of the speciation and the biological transfer of Pu in aquatic ecosystems.

The technique of diffusive gradients in thin films (DGT) is proposed for speciation studies of plutonium. This protocol describes diffusion experiments probing the behavior of Pu(IV) and Pu(V) in presence of organic matter. DGTs deployed in a karstic spring allow assessment of the bioavailability of Pu.

环境钚的薄膜弥散形态和生物利用度测量

The biological uptake of plutonium (Pu) in aquatic ecosystems is of particular concern since it is an alpha-particle emitter with long half-life which can ...

In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis

Micro-analytical techniques based on chemical element imaging enable the localization and quantification of chemical composition at the cellular level. They offer new possibilities for the characterization of living systems and are particularly appropriate for detecting, localizing and quantifying the presence of metal oxide nanoparticles both in biological specimens and the environment. Indeed, these techniques all meet relevant requirements in terms of (i) sensitivity (from 1 up to 10 &#181;g.g-1 of dry mass), (ii) micrometer range spatial resolution, and (iii) multi-element detection. Given these characteristics, microbeam chemical element imaging can powerfully complement routine imaging techniques such as optical and fluorescence microscopy. This protocol describes how to perform a nuclear microprobe analysis on cultured cells (U2OS) exposed to titanium dioxide nanoparticles. Cells must grow on and be exposed directly in a specially designed sample holder used on the optical microscope and in the nuclear microprobe analysis stages. Plunge-freeze cryogenic fixation of the samples preserves both the cellular organization and the chemical element distribution. Simultaneous nuclear microprobe analysis (scanning transmission ion microscopy, Rutherford backscattering spectrometry and particle induced X-ray emission) performed on the sample provides information about the cellular density, the local distribution of the chemical elements, as well as the cellular content of nanoparticles. There is a growing need for such analytical tools within biology, especially in the emerging context of Nanotoxicology and Nanomedicine for which our comprehension of the interactions between nanoparticles and biological samples must be deepened. In particular, as nuclear microprobe analysis does not require nanoparticles to be labelled, nanoparticle abundances are quantifiable down to the individual cell level in a cell population, independently of their surface state.

In Situ Detection and Single Cell Quantification of Metal Oxide Nanoparticles Using Nuclear Microprobe Analysis

We describe a procedure for the detection of chemical elements present in situ in human cells as well as their in vitro quantification. The method is well-suited to any cell type and is particularly useful for quantitative chemical analyses in single cells following in vitro metal oxide nanoparticles exposure.

原位金属氧化物纳米粒子的核探针检测与单细胞定量研究

Micro-analytical techniques based on chemical element imaging enable the localization and quantification of chemical composition at the cellular level. ...

化学

Two-way Valorization of Blast Furnace Slag: Synthesis of Precipitated Calcium Carbonate and Zeolitic Heavy Metal Adsorbent

The aim of this work is to present a zero-waste process for storing CO2 in a stable and benign mineral form while producing zeolitic minerals with sufficient heavy metal adsorption capacity. To this end, blast furnace slag, a residue from iron-making, is utilized as the starting material. Calcium is selectively extracted from the slag by leaching with acetic acid (2 M CH3COOH) as the extraction agent. The filtered leachate is subsequently physico-chemically purified and then carbonated to form precipitated calcium carbonate (PCC) of high purity (&lt;2 wt% non-calcium impurities, according to ICP-MS analysis). Sodium hydroxide is added to neutralize the regenerated acetate. The morphological properties of the resulting calcitic PCC are tuned for its potential application as a filler in papermaking. In parallel, the residual solids from the extraction stage are subjected to hydrothermal conversion in a caustic solution (2 M NaOH) that leads to the predominant formation of a particular zeolitic mineral phase (detected by XRD), namely analcime (NaAlSi2O6&#8729;H2O). Based on its ability to adsorb Ni2+, as reported from batch adsorption experiments and ICP-OES analysis, this product can potentially be used in wastewater treatment or for environmental remediation applications.

A protocol for the parallel production of precipitated calcium carbonate and zeolitic material from blast furnace slag via mineral carbonation and alkaline hydrothermal conversion, respectively, is presented. The performance of the zeolitic material towards nickel adsorption is tested.

高炉矿渣的双向稳定物价:沉淀碳酸钙和沸石重金属吸附剂的合成

The aim of this work is to present a zero-waste process for storing CO2 in a stable and benign mineral form while producing zeolitic minerals with ...

Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

Uranium has been shown to interfere with bone physiology and it is well established that this metal accumulates in bone. However, little is known about the effect of natural uranium on the behavior of bone cells. In particular, the impact of uranium on osteoclasts, the cells responsible for the resorption of the bone matrix, is not documented. To investigate this issue, we have established a new protocol using uranyl acetate as a source of natural uranium and the murine RAW 264.7 cell line as a model of osteoclast precursors. Herein, we detailed all the assays required to test uranium cytotoxicity on osteoclast precursors and to evaluate its impact on the osteoclastogenesis and on the resorbing function of mature osteoclasts. The conditions we have developed, in particular for the preparation of uranyl-containing culture media and for the seeding of RAW 264.7 cells allow to obtain reliable and highly reproductive results. Moreover, we have optimized the use of software tools to facilitate the analysis of various parameters such as the size of osteoclasts or the percentage of resorbed matrix.

Methods for Analyzing the Impacts of Natural Uranium on In Vitro Osteoclastogenesis

Uranium is known to affect bone metabolism. Here, we present a protocol aimed at investigating the effect of natural uranium exposure on the viability, the differentiation, and the function of osteoclasts, the cells in charge of bone resorption.

天然铀对体外破影响的分析方法

Uranium has been shown to interfere with bone physiology and it is well established that this metal accumulates in bone. However, little is known about ...

发育生物学

U2O5 Film Preparation via UO2 Deposition by Direct Current Sputtering and Successive Oxidation and Reduction with Atomic Oxygen and Atomic Hydrogen

We describe a method to produce U2O5 films in situ using the Labstation, a modular machine developed at JRC Karlsruhe. The Labstation, an essential part of the Properties of Actinides under Extreme Conditions laboratory (PAMEC), allows the preparation of films and studies of sample surfaces using surface analytical techniques such as X-ray and ultra-violet photoemission spectroscopy (XPS and UPS, respectively). All studies are made in situ, and the films, transferred under ultra-high vacuum from their preparation to an analyses chamber, are never in contact with the atmosphere. Initially, a film of UO2 is prepared by direct current (DC) sputter deposition on a gold (Au) foil then oxidized by atomic oxygen to produce a UO3 film. This latter is then reduced with atomic hydrogen to U2O5. Analyses are performed after each step involving oxidation and reduction, using high-resolution photoelectron spectroscopy to examine the oxidation state of uranium. Indeed, the oxidation and reduction times and corresponding temperature of the substrate during this process have severe effects on the resulting oxidation state of the uranium. Stopping the reduction of UO3 to U2O5 with single U(V) is quite challenging; first, uranium-oxygen systems exist in numerous intermediate phases. Second, differentiation of the oxidation states of uranium is mainly based on satellite peaks, whose intensity peaks are weak. Also, experimenters should be aware that X-ray spectroscopy (XPS) is a technique with an atomic sensitivity of 1% to 5%. Thus, it is important to obtain a complete picture of the uranium oxidation state with the entire spectra obtained on U4f, O1s, and the valence band (VB). Programs used in the Labstation include a linear transfer program developed by an outside company (see Table of Materials) as well as data acquisition and sputter source programs, both developed in-house.

U2O5 Film Preparation via UO2 Deposition by Direct Current Sputtering and Successive Oxidation and Reduction with Atomic Oxygen and Atomic Hydrogen

This protocol presents the preparation of U2O5 thin films obtained in situ under ultra-high vacuum. The process involves oxidation and reduction of UO2 films with atomic oxygen and atomic hydrogen, respectively.

利用uo2沉积制备 u2o5 薄膜, 利用原子氧和原子氢连续氧化还原

We describe a method to produce U2O5 films in situ using the Labstation, a modular machine developed at JRC Karlsruhe. The Labstation, an essential part ...

Separation of Uranium and Thorium for 230Th-U Dating of Submarine Hydrothermal Sulfides

The age of a submarine hydrothermal sulfide is a significant index for estimating the size of hydrothermal ore deposits. Uranium and thorium isotopes in the samples can be separated for 230Th-U dating. This article presents a method to purify and separate U and Th isotopes in submarine hydrothermal sulfide samples. Following this technique, the separated U and Th fractions can meet measuring requirements by multi-collector inductively coupled plasma mass spectrometry (MC-ICPMS). The age of the hydrothermal sulfide sample can be calculated by measuring the present-day activity ratios of 230Th/238U and 234U/238U. A super clean room is necessary for this experiment. Cleaned regents and supplies are used to reduce the contamination during the sample processes. Balance, hotplate, and centrifuge are also used. The sulfide sample is powdered for analysis and less than 0.2 g sample is used. Briefly, the sample is weighed, dissolved, added to 229Th-233U-236U double spike solution, Fe co-precipitated, and separated on an anion-exchange resin extraction column. Approximately 50 ng U is consumed for 230Th-U dating of sulfides sample by MC-ICPMS.

Separation of Uranium and Thorium for 230Th-U Dating of Submarine Hydrothermal Sulfides

The protocol describes a method to purify and separate the U and Th nuclide in submarine hydrothermal sulfide sample with Fe co-precipitation and extraction chromatography for 230Th-U disequilibrium dating.

海底热液硫化物230Th-U测定铀和铀的分离

The age of a submarine hydrothermal sulfide is a significant index for estimating the size of hydrothermal ore deposits. Uranium and thorium isotopes in ...

Accumulation and Analysis of Cuprous Ions in a Copper Sulfate Plating Solution

Knowledge of the behavior of cuprous ions (monovalent copper ion: Cu(I)) in a copper sulfate plating bath is important for improving the plating process. We successfully developed a method to quantitatively and easily measure Cu(I) in a plating solution and used it for evaluation of the solution. In this paper, a quantitative absorption spectrum measurement and a time-resolved injection measurement of Cu(I) concentrations by a color reaction are described. This procedure is effective as a method to reproduce and elucidate the phenomenon occurring in the plating bath in the laboratory. First, the formation and accumulation process of Cu(I) in solution by electrolysis of a plating solution is shown. The amount of Cu(I) in the solution is increased by electrolysis at higher current values &#8203;&#8203;than the usual plating process. For the determination of Cu(I), BCS (bathocuproinedisulfonic acid, disodium salt), a reagent that selectively reacts with Cu(I), is used. The concentration of Cu(I) can be calculated from the absorbance of the Cu(I)-BCS complex. Next, the time measurement of the color reaction is described. The color reaction curve of Cu(I) and BCS measured by the injection method can be decomposed into an instantaneous component and a delay component. By analysis of these components, the holding structure of Cu(I) can be clarified, and this information is important when predicting the quality of the plating film to be produced. This method is used to facilitate the evaluation of the plating bath in the production line.

Here, accumulation of cuprous ions in a copper sulfate plating solution in a model experiment and an analysis based on quantitative measurements are described. This experiment reproduces the accumulation process of cuprous ions in the plating bath.

硫酸铜镀液中铜离子的积累与分析

Knowledge of the behavior of cuprous ions (monovalent copper ion: Cu(I)) in a copper sulfate plating bath is important for improving the plating process. ...

Kupffer Cell Isolation for Nanoparticle Toxicity Testing

The large majority of in vitro nanotoxicological studies have used immortalized cell lines for their practicality. However, results from nanoparticle toxicity testing in immortalized cell lines or primary cells have shown discrepancies, highlighting the need to extend the use of primary cells for in vitro assays. This protocol describes the isolation of mouse liver macrophages, named Kupffer cells, and their use to study nanoparticle toxicity. Kupffer cells are the most abundant macrophage population in the body and constitute part of the reticulo-endothelial system (RES), responsible for the capture of circulating nanoparticles. The Kupffer cell isolation method reported here is based on a 2-step perfusion method followed by purification on density gradient. The method, based on collagenase digestion and density centrifugation, is adapted from the original protocol developed by Smedsr&#248;d et al. designed for rat liver cell isolation and provides high yield (up to 14 x 106 cells per mouse) and high purity (&#62;95%) of Kupffer cells. This isolation method does not require sophisticated or expensive equipment and therefore represents an ideal compromise between complexity and cell yield. The use of heavier mice (35-45 g) improves the yield of the isolation method but also facilitates remarkably the procedure of portal vein cannulation. The toxicity of functionalized carbon nanotubes f-CNTs was measured in this model by the modified LDH assay. This method assesses cell viability by measuring the lack of structural integrity of Kupffer cell membrane after incubation with f-CNTs. Toxicity induced by f-CNTs can be measured consistently using this assay, highlighting that isolated Kupffer cells are useful for nanoparticle toxicity testing. The overall understanding of nanotoxicology could benefit from such models, making the nanoparticle selection for clinical translation more efficient.

Liver macrophages, named Kupffer cells, are responsible for the capture of circulating nanoparticles. We describe here a method, of high cell purity and yield, for Kupffer cell isolation. The modified LDH assay is used here to measure the toxicity induced by carbon nanotubes in Kupffer cells.

枯否细胞分离的纳米粒子毒性测试

The large majority of in vitro nanotoxicological studies have used immortalized cell lines for their practicality. However, results from nanoparticle ...

去除微量元素的氧化铜纳米粉体制备铀

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