表确认和资金来源。 NIH-ES015022，ES018274（TRN） NSF-1003907合作协议（VCM）

全身纳米粒子吸入暴露一步一步的操作过程进行了描述如下。 注 ：1）步骤1和3应该在通风橱内进行; 2）操作人员必须穿戴适当的个人防护装备（口罩，护目镜和橡胶手套）。 1。空调的TiO 2纳米干粉<ol><li>将纳米TiO 2粉体在不透明的容器中。 </li><li>保持容器盖打开。 </li><li>用于调节至少24小时，将容器放置在干燥器中干燥。 </li></ol> 2。数据采集​​和控制系统，SMPS和ELPI和所有传感器热身<ol><li>打开空气监控和数据采集系统和气溶胶监测SMPS（TSI公司肖尔维尤，明尼苏达州）ELPI（Dekati，坦佩雷，芬兰），电源开关，暖系统至少1小时。 </li><li>打开电源所有传感器的开关温暖起来至少1小时。 </li></ol> 3。载入TiO 2纳米的干粉成气溶胶发生器<ol><li>气溶胶发生器打开汽缸盖，并更换过滤器中的气溶胶发生器。注：一个气溶胶发生器有一个缸。要使用的气溶胶发生器的数量取决于所需的曝光室中的粒子的质量浓度。 </li><li>称重〜4克纳米TiO 2粉体，并加载它们在每个气缸。 </li><li>更换气缸盖。 </li><li>所有区域应怀疑二氧化钛污染湿抹。 </li></ol> 4。气溶胶发生器连接到吸入暴露商会<ol><li>连接气溶胶发生器通过歧管旋风分离器是吸入暴露室的入口（TSE Systems GmbH公司在德国Bad Homburg）的所有出水口。 </li><li>连接压缩空气管文丘里分散在气溶胶发生器。 </li></ol> 5。连接空气监测和气溶胶采样入口吸入暴露室<ol><li>连接的温度和相对湿度（RH），压力，O 2和CO 2传感器由TSE系统提供测试大气监测端口吸入暴露室。 </li><li>气溶胶取样口上的吸入暴露室的气溶胶稀释器的入口连接，然后连接它的出口ELPI的入口。 </li><li>将开关电源连接的一个上的吸入暴露室的气溶胶取样口。 </li><li>入口的粒子浓度监测（TSE系统）连接到一个在曝光室的气溶胶取样口。 </li><li>称重PTFE膜过滤器（P / N 66149），颇尔公司，密歇根州安阿伯和加载滤入一个不锈钢过滤器固定架（TOX产品，莫里亚蒂NM）。 </li><li>连接的入口的不锈钢过滤器保持器与一个预先称重的过滤器的一个上的吸入暴露室的气溶胶取样口，并连接到取样泵的出口。 </li></ol> 6。激活数据采集系统<ol><li>激活ELPI数据采集软件，ELPIVI，检查设置参数，打开冲洗泵〜5分钟，然后归零ELPI。记录预曝光浓度。 </li><li>激活SMPS的数据采集软件。记录预曝光浓度。 </li><li>激活软件， 达戈 （TSE），用于监视和控制空气流速，温度和相对湿度腔的压力，温度和湿度，O 2和CO 2。 </li></ol> 7。加载实验动物吸入暴露室<ol><li>称量实验动物。 </li><li>实验动物和网箱标记可以使动物放回曝光后，在同一个笼子里，如果娘家DED。 </li><li>吸入暴露室的门打开，并加载到有线笼实验动物。 </li><li>可将水提供动物。 </li><li>关闭并固定吸入暴露室的门。 </li><li>经常观察动物通过曝光室观察窗遇险的迹象。动物应该放松，行为正常。如果快速停止曝光/呼吸困难，外观异常，姿势异常或行动不便观察。取出的动物，他们返回他们原来的笼子，请联系主治兽医和/或采取适当的机构动物护理和使用委员会的程序。 </li></ol> 注：执行步骤8.7，8.8和8.17时，操作人员必须穿戴个人防护装备。 8。纳米气溶胶使小动物<ol><li>打开排气真空泵吸入暴露室。 </li><li>运行数据采集软件的Daco）曝光室提供过滤的干燥空气，B）控制曝光室中的压力，和c）收集暴露环境中的数据，如压力，温度，相对湿度，O 2和CO 2。 </li><li>建立微负压室压力（设定点= -0.2毫巴）。 </li><li>气溶胶发生器打开。 </li><li>运行ELPI和SMPS数据采集软件的连续监测颗粒大小和相对质量浓度吸入暴露室。 </li><li>当气溶胶浓度是稳定的， 即到达高原ELPI显示器上的浓度分布曲线（通常情况下：后的气溶胶发生器是在操作的，这需要20分钟），设置采样时间（例如，1小时），并打开气溶胶采样纳米粒子过滤器，泵，收集有代表性的样品。 </li><li>一旦采样时间到达，取出过滤器堵塞SAmpling港口用橡胶塞，以防止测试材料曝光室逃逸。 </li><li>称重的过滤器，并计算出在如上所述的曝光室的平均质量浓度。 </li><li>如果平均浓度是关闭的目标浓度时，手动调节发电机中的气流，以确保达到目标浓度。 </li><li>粒子计算为D = C x垂直米 XTX F R，其中D =剂量，C =平均质量浓度的试验材料，V 米 =每分钟通气量， 吨 =暴露时间，F R =小部分的材料沉积在动物的肺沉积或吸收。 </li><li>更换过滤器在过滤干净，预加权滤波器持有人，并重复步骤8.6和8.8。 </li><li>根据真实的质量浓度在曝光室和有针对性的颗粒沉积在动物肺，估计余下的EXP，T 保持 osure时间=（ð 针对性 -D）/（C X V M X F R），其中t 仍然保持暴露时间，D 有针对性的 =有针对性剂量，C =测试材料的平均质量浓度，V M =每分钟通气量，F R =馏分材料沉积或吸收。 </li><li> 当 t达到保持关闭气溶胶发生器。 </li><li>从曝光室之前除去动物，冲洗吸入的曝光室用过滤后的空气，直到在显示屏中显示的粒子浓度接近预曝光在室中的颗粒浓度。 </li><li>关闭室排气真空泵。 </li><li>停止， 达戈数据采集软件。 </li><li>曝光后，观察动物是否能够正常呼吸和行为，并没有其他的文件学习并发症前北京时间。如果鼻腔分泌物，呼吸窘迫或任何其他动物福利并发症的观察，联系主治兽医和/或采取适当的机构动物护理和使用委员会的程序。 </li><li>停止ELPI和SMPS数据采集软件的。 </li></ol> 9。创建测试报告 9.1测试条件包括： <ol><li>此测试中使用的烟雾生成系统，其运行参数的描述。 </li><li>包括设计，类型，尺寸和其运行参数，在曝光过程中使用的曝光装置的说明。 </li><li>设备用于测量温度，湿度，颗粒尺寸，和实际浓度。 </li><li>排出的空气和住房动物在试验室中使用时的方法处理。 </li></ol> 9.2曝光大气数据包括<ol><li>通过吸入设备的气流速率。 </li><li>温度和湿度空气。 </li><li>实际（分析或重量）浓度的气溶胶采样区附近的动物笼子。 </li><li>的粒度分布，计算出的计数中位数气体动力学直径和几何标准偏差。 </li><li>解释为什么想要室浓度和/或颗粒大小无法实现（如适用），及所作的努力遵守这些方面的指引。 </li></ol> 9.3其他<ol><li>应保持微负压在室内吸入含设施，防止逃避吸入暴露实验室测试材料。 </li><li>每天清洁曝光室的动物废弃物，以消除影响。 </li><li> ELPI，SMPS和其它的仪器应进行清洗和校准的基础上的用户手册。 </li></ol>

<ol>
	<li>Bide, R. W., Armour, S. J., Yee, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Allometric+respiration/body+mass+data+for+animals+to+be+used+for+estimates+of+inhalation+toxicity+to+young+adult+humans.">Allometric respiration/body mass data for animals to be used for estimates of inhalation toxicity to young adult humans.</a> J. Appl. Toxicol. 20 (4), 273-290 (2000).</li><li>Guyton, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analysis+of+respiratory+patterns+in+laboratory+animals.">Analysis of respiratory patterns in laboratory animals.</a> Am. J. Physiol. 150, 70-77 (1947).</li><li>Knuckles, T. L., Yi, J., Frazer, D. G., Leonard, H. D., Chen, B. T., Castranova, V., Nurkiewicz, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticle+inhalation+alters+systemic+arteriolar+vasoreactivity+through+sympathetic+and+cyclooxygenase-mediated+pathways.">Nanoparticle inhalation alters systemic arteriolar vasoreactivity through sympathetic and cyclooxygenase-mediated pathways.</a> Nanotoxicology. , 1-12 (2011).</li><li>Pauluhn, J., Mohr, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Repeated+4-week+inhalation+exposure+of+rats:+effect+of+low-,+intermediate,+and+high-humidity+chamber+atmosphere.">Repeated 4-week inhalation exposure of rats: effect of low-, intermediate, and high-humidity chamber atmosphere.</a> Exp. Toxic Pathol. , 178-187 (1999).</li><li>Schmoll, L. H., Elzey, S., Grassian, V. H., O'Shaughnessy, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticle+aerosol+generation+methods+from+bulk+powders+for+inhalation+exposure+studies.">Nanoparticle aerosol generation methods from bulk powders for inhalation exposure studies.</a> Nanotoxicology. 3, 265-275 (2009).</li><li>To, D., Yin, X., Sundaresan, S., Dave, R. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deagglomeration+of+nano-particle+aggregates+via+rapid+expansion+of+high+pressure+suspensions.">Deagglomeration of nano-particle aggregates via rapid expansion of high pressure suspensions.</a> AIChE J. 55 (11), 2756-3032 (2009).</li><li>U.S. Environmental Protection Agency (US EPA). <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Health+effects+test+guidelines:+OPPTS.,+870.1300.+Acute+inhalation+toxicity.">Health effects test guidelines: OPPTS., 870.1300. Acute inhalation toxicity.</a> EPA. , 712-C-98-193 (1998).</li><li>Wong, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+feedback+control+of+an+inhalation+exposure+system+with+discrete+sampling+intervals:+testing,+performance,+and+modeling.">Automated feedback control of an inhalation exposure system with discrete sampling intervals: testing, performance, and modeling.</a> Inhal. Toxicol. 15, 729-743 (2003).</li><li>Wong, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inhalation+Exposure+Systems:+Design,+Methods+and+Operation.">Inhalation Exposure Systems: Design, Methods and Operation.</a> Toxicologic Pathology. 35, 3-14 (2007).</li><li>Yi, J., Nurkiewicz, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanoparticle+Aerosol+Generator.">Nanoparticle Aerosol Generator.</a> US patent. , (2011).</li></ol>

本报告中的调查结果和结论的作者，并不一定代表职业安全和健康研究所的意见。提及任何公司名称或产品并不意味着赞同通过NIOSH，也不意味着替代产品是不可用，或者无法被取代适当的评估后。

我们已经组建，并在这里描述一个全身纳米粒子雾化吸入曝光系统。该系统的功能进行了验证与国家的最先进的纳米粒子气溶胶的表征技术。该吸入暴露系统可以采用一种新型的纳米粒子的气溶胶的产生系统，其特征在于，控制和均匀的纳米粒子的气溶胶测试气氛相对一致的温度，湿度，空气流量和氧含量为实验动物。曝光系统是最有效的大量的动物，或长期的研究。在这种大的全身室，实验动?...

<table border="1">
		<tbody>
		 <tr>
			<td>Name of Reagent/Material</td>
			<td>Company</td>
			<td>Catalog Number</td>
			<td>Comments</td>
		 </tr>
		 <tr>
			<td>Inhalation exposure system</td>
			<td>TSE Systems GmbH, Bad Homburg, Germany</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Air monitoring system</td>
			<td>TSE Systems GmbH, Bad Homburg, Germany</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Titanium dioxide Aeroxide P25</td>
			<td>Evonik, Germany</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Scanning mobility particle sizer-3936L75</td>
			<td>TSI Inc., Shoreview, MN</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Electric low pressure impactor, Standard 10 LPM</td>
			<td>Dekati, Tampere, Finland</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Ultra Micro Balance, XP2U</td>
			<td>METTLER TOLEDO, Switzerland</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Field Emission Scanning Electron Microscope-S-4800</td>
			<td>Hitachi, Japan</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Energy dispersive X-ray analysis </td>
			<td>Princeton Gamma-Tech, Rocky Hill, N.J.</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>Nuclepore polycarbonate filters </td>
			<td>Whatman, Clinton, PA</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>PTFE membrane filters </td>
			<td>Pall corporation, Ann Arbor, Michigan </td>
			<td></td>
			<td></td>
		 </tr>
		</tbody>
 </table>

whole-body nanoparticle aerosol inhalation exposures

吸入与aerosolizable工程纳米材料（ENM）工作的个人，是最有可能的暴露途径。适当地进行纳米粒子吸入毒理学研究，在一个腔室壳体中的气溶胶实验动物必须具有：1）为整个曝光期间所需的电平保持在一个稳定的浓度; 2）均匀的组合物不含任何污染物，以及3）一个稳定的粒度分布，几何平均直径小于200 nm和几何标准偏差σ&lt;2.5 5 克 。含有纳米粒子的气溶胶产生相当具有挑战性的，因为纳米粒子易结块。这主要是由于非常强的粒子间力而形成的大的分形结构，在数十或数百个大小6微米，这是很难被打破。几种常见的气溶胶发生器，雾化器，流化床，文丘里吸气和怀特粉尘饲料，重新测试，但是，没有人能够产生纳米粒子气溶胶满足所有标准5。 一整体纳米粒子的气溶胶吸入暴露系统制备纳米TiO 2的吸入毒理学研究，验证和使用。关键部件：1）新型纳米TiO 2的气溶胶发生器; 2）0.5米3全身吸入暴露室，以及3）监视和控制系统。纳米TiO 2材料产生的干散纳米TiO 2粉末（一次粒径为21 nm，堆积密度为3.8克/厘米3）的气溶胶被交付到曝光室90 LPM（10.8空中更改/小时）的流速。连续测量的粒径分布和质量浓度分布与流动性的扫描粒度仪（SMPS），和一个电动的低压冲击器（ELPI）。重量法的气溶胶质量浓度（C）进行了验证（ 毫克/ 立方米 ）。质量（M）所收集的粒子被确定为M =（M-M 后前 ）， 前 ，其中M和M 邮政群众的过滤器前后采样（ 毫克 ）。的质量浓度计算为：C = M /（Q *吨），其中Q为采样流量（ 米3 /分），t为采样时间（ 分钟 ）。腔室的压力，温度，相对湿度（RH），O 2和CO 2浓度的连续监测和控制。纳米TiO 2气溶胶核孔过滤器上收集，用扫描电子显微镜（SEM）和能量色散X射线分析（EDX）分析。 综上所述，我们报告说，纳米粒子气溶胶的生成和交付我们的曝光室有：1）稳定的质量浓度; 2）同质组成无污染物; 3）与计数中位数aerody的稳定的粒度分布动力学直径157nm的气溶胶的产生过程中。该系统可靠，并多次创建测试环境，模拟职业，环境或国内安宁气溶胶风险。

一种吸入暴露研究通常涉及保持在已知和恒定的测试环境中的实验动物实验动物暴露至规定浓度的试验物质8,9。 图1中所示的全身纳米粒子吸入暴露系统。全身室是一个动态的流量基础上操作，那里有一个连续90 LPM气流通过室。这股气流10.8空气更改/小时，超过空气交换的最小数量由美国环境保护署 （10.0）急性吸入暴露7。 3级空气过滤系统，包括一个凝聚式过?...

Watch this Scientific Journal Video about Whole-Body Nanoparticle Aerosol Inhalation Exposures at JoVE.com

Whole-Body Nanoparticle Aerosol Inhalation Exposures

一整体纳米粒子的气溶胶吸入暴露设施建造纳米级钛白粉（二氧化钛<sub&gt; 2</sub&gt;）吸入毒理学研究。此系统提供了纳米TiO<sub&gt; 2</sub烟雾测试环境，有：1）稳定的质量浓度，2）均匀的组合物不含任何污染物，以及3）稳定的气溶胶的产生过程中的颗粒尺寸分布。

全身纳米气溶胶吸入暴露

Inhalation is the most likely exposure route for individuals working with aerosolizable engineered nano-materials (ENM). To properly perform nanoparticle ...

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JoVE Journal

Biology

15.7K 次观看.  West Virginia University. 一整体纳米粒子的气溶胶吸入暴露设施建造纳米级钛白粉（二氧化钛 2）吸入毒理学研究。此系统提供了纳米TiO 2

视频: Whole-Body Nanoparticle Aerosol Inhalation Exposures

Measuring Respiratory Function in Mice Using Unrestrained Whole-body Plethysmography

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative assessment of lung function in rodent models is an important tool in the development of future therapies. Commonly used techniques for assessing respiratory function including invasive plethysmography and forced oscillation. While these techniques provide valuable information, data collection can be fraught with artefacts and experimental variability due to the need for anesthesia and/or invasive instrumentation of the animal. In contrast, unrestrained whole-body plethysmography (UWBP) offers a precise, non-invasive, quantitative way by which to analyze respiratory parameters. This technique avoids the use of anesthesia and restraints, which is common to traditional plethysmography techniques. This video will demonstrate the UWBP procedure including the equipment set up, calibration and lung function recording. It will explain how to analyze the collected data, as well as identify experimental outliers and artefacts that results from animal movement. The respiratory parameters obtained using this technique include tidal volume, minute volume, inspiratory duty cycle, inspiratory flow rate and the ratio of inspiration time to expiration time. UWBP does not rely on specialized skills and is inexpensive to perform. A key feature of UWBP, and most appealing to potential users, is the ability to perform repeated measures of lung function on the same animal.

The assessment of respiratory physiology has traditionally relied upon techniques, which require restraint or sedation of the animal. Unrestrained whole-body plethysmography, however, provides precise, non-invasive, quantitative analysis of respiratory physiology in animal models. In addition, the technique allows repeated respiratory assessment of mice allowing for longitudinal studies.

用奔放的整体体积描记测量呼吸功能的影响

Respiratory dysfunction is one of the leading causes of morbidity and mortality in the world and the rates of mortality continue to rise. Quantitative ...

科研

生物学

Chitosan/Interfering RNA Nanoparticle Mediated Gene Silencing in Disease Vector Mosquito Larvae

Vector mosquitoes inflict more human suffering than any other organism&#8212;and kill more than one million people each year. The mosquito genome projects facilitated research in new facets of mosquito biology, including functional genetic studies in the primary African malaria vector Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti. RNA interference- (RNAi-) mediated gene silencing has been used to target genes of interest in both of these disease vector mosquito species. Here, we describe a procedure for preparation of chitosan/interfering RNA nanoparticles that are combined with food and ingested by larvae. This technically straightforward, high-throughput, and relatively inexpensive methodology, which is compatible with long double stranded RNA (dsRNA) or small interfering RNA (siRNA) molecules, has been used for the successful knockdown of a number of different genes in A. gambiae and A. aegypti larvae. Following larval feedings, knockdown, which is verified through qRT-PCR or in situ hybridization, can persist at least through the late pupal stage. This methodology may be applicable to a wide variety of mosquito and other insect species, including agricultural pests, as well as other non-model organisms. In addition to its utility in the research laboratory, in the future, chitosan, an inexpensive, non-toxic and biodegradable polymer, could potentially be utilized in the field.

Here we describe a procedure for inhibiting gene function in disease vector mosquitoes through the use of chitosan/interfering RNA nanoparticles that are ingested by larvae.

壳聚糖/干扰RNA纳米介导的基因沉默的疾病病媒蚊幼虫

Vector mosquitoes inflict more human suffering than any other organism&#8212;and kill more than one million people each year. The mosquito genome projects ...

A Method for Selecting Structure-switching Aptamers Applied to a Colorimetric Gold Nanoparticle Assay

Small molecules provide rich targets for biosensing applications due to their physiological implications as biomarkers of various aspects of human health and performance. Nucleic acid aptamers have been increasingly applied as recognition elements on biosensor platforms, but selecting aptamers toward small molecule targets requires special design considerations. This work describes modification and critical steps of a method designed to select structure-switching aptamers to small molecule targets. Binding sequences from a DNA library hybridized to complementary DNA capture probes on magnetic beads are separated from nonbinders via a target-induced change in conformation. This method is advantageous because sequences binding the support matrix (beads) will not be further amplified, and it does not require immobilization of the target molecule. However, the melting temperature of the capture probe and library is kept at or slightly above RT, such that sequences that dehybridize based on thermodynamics will also be present in the supernatant solution. This effectively limits the partitioning efficiency (ability to separate target binding sequences from nonbinders), and therefore many selection rounds will be required to remove background sequences. The reported method differs from previous structure-switching aptamer selections due to implementation of negative selection steps, simplified enrichment monitoring, and extension of the length of the capture probe following selection enrichment to provide enhanced stringency. The selected structure-switching aptamers are advantageous in a gold nanoparticle assay platform that reports the presence of a target molecule by the conformational change of the aptamer. The gold nanoparticle assay was applied because it provides a simple, rapid colorimetric readout that is beneficial in a clinical or deployed environment. Design and optimization considerations are presented for the assay as proof-of-principle work in buffer to provide a foundation for further extension of the work toward small molecule biosensing in physiological fluids.

A protocol is provided to select structure-switching aptamers for small molecule targets based on a tunable stringency magnetic bead selection method. Aptamers selected with structure-switching properties are beneficial for assays that require a conformational change to signal the presence of a target, such as the described gold nanoparticle assay.

适用于比色金纳米粒子检测方法的选择结构切换适配方法

Small molecules provide rich targets for biosensing applications due to their physiological implications as biomarkers of various aspects of human health ...

Design and Development of Aptamer&#8211;Gold Nanoparticle Based Colorimetric Assays for In-the-field Applications

The design and development of an aptamer&#8211;gold nanoparticle (AuNP) colorimetric assay for the detection of small molecules for in-the-field applications was examined. Target selective AuNP based color assays have been developed in controlled proof-of-concept laboratory settings. However, these schemes have not been exerted to a point of failure to determine their practical use beyond laboratory settings. This work describes a generic approach to design, develop, and troubleshoot an aptamer-AuNP colorimetric assay for small molecule analytes and using the assay for in-the-field settings. The assay is advantageous because adsorbed aptamers passivate the nanoparticle surfaces and provide a means to reduce and eliminate false positive responses to non-target analytes. Transitioning this system to practical uses required defining not only the shelf-life of the aptamer-AuNP assay, but establishing methods and procedures for extending the long-term storage capabilities. Also, one of the recognized concerns with colorimetric readout is the burden placed on analysts to accurately identify often subtle changes in color. To lessen the responsibility on analysts in the field, a color analysis protocol was designed to perform the color identification duties without the need for performing this task on laboratory grade equipment. The method for creating and testing the data analysis protocol is described. However to understand and influence the design of adsorbed aptamer assays, the interactions associated with the aptamer, target, and AuNPs require further study. The knowledge gained could lead to tailoring aptamers for improved functionality.

The design and development of an aptamer&#8211;gold nanoparticle colorimetric assay for the detection of small molecules for in-the-field applications was examined. Additionally, a smart-device colorimetric application (app) was validated and long-term storage of the assay was established for use in the field.

设计和适体 - 金纳米粒子基于比色法的开发在这领域的应用

The design and development of an aptamer&#8211;gold nanoparticle (AuNP) colorimetric assay for the detection of small molecules for in-the-field ...

An Air-liquid Interface Bronchial Epithelial Model for Realistic, Repeated Inhalation Exposure to Airborne Particles for Toxicity Testing

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic exposure conditions. This puts specific demands on the cell culture models. Many cell types are negatively affected by exposure to air (e.g., drying out) and only remain viable for a few days. This limits the exposure conditions that can be used in these models: usually relatively high concentrations are applied as a cloud (i.e., droplets containing particles, which settle down rapidly) within a short period of time. Such experimental conditions do not reflect realistic long-term exposure to low concentrations of particles. To overcome these limitations the use of a human bronchial epithelial cell line, Calu-3 was investigated. These cells can be cultured at ALI conditions for several weeks while retaining a healthy morphology and a stable monolayer with tight junctions. In addition, this bronchial model is suitable for testing the effects of repeated exposures to low, realistic concentrations of airborne particles using an ALI exposure system. This system uses a continuous airflow in contrast to other ALI exposure systems that use a single nebulization producing a cloud. Therefore, the continuous flow system is suitable for repeated and prolonged exposure to airborne particles while continuously monitoring the particle characteristics, exposure concentration, and delivered dose. Taken together, this bronchial model, in combination with the continuous flow exposure system, is able to mimic realistic, repeated inhalation exposure conditions that can be used for toxicity testing.

Described is the cell culture and exposure method of an in vitro bronchial model for realistic, repeated inhalation exposure to particles for toxicity testing.

用于真实、反复吸入空气颗粒以进行毒性测试的气液接口支气管上皮模型

For toxicity testing of airborne particles, air-liquid interface (ALI) exposure systems have been developed for in vitro tests in order to mimic realistic ...

Dosimetry for Cell Irradiation using Orthovoltage (40-300 kV) X-Ray Facilities

The importance of dosimetry protocols and standards for radiobiological studies is self-evident. Several protocols have been proposed for dose determination using low energy X-ray facilities, but depending on the irradiation configurations, samples, materials or beam quality, it is sometimes difficult to know which protocol is the most appropriate to employ. We, therefore, propose a dosimetry protocol for cell irradiations using low energy X-ray facility. The aim of this method is to perform the dose estimation at the level of the cell monolayer to make it as close as possible to real cell irradiation conditions. The different steps of the protocol are as follows: determination of the irradiation parameters (high voltage, intensity, cell container etc.), determination of the beam quality index (high voltage-half value layer couple), dose rate measurement with ionization chamber calibrated in air kerma conditions, quantification of the attenuation and scattering of the cell culture medium with EBT3 radiochromic films, and determination of the dose rate at the cellular level. This methodology must be performed for each new cell irradiation configuration as the modification of only one parameter can strongly impact the real dose deposition at the level of the cell monolayer, particularly involving low energy X-rays.

Dosimetry for Cell Irradiation using Orthovoltage 40-300 kV X-Ray Facilities

This document describes a new dosimetry protocol for cell irradiations using low energy X-ray equipment. Measurements are performed in conditions simulating real cell irradiation conditions as much as possible.

使用矫形器（40-300 kV）X射线设施进行细胞辐照的测定

The importance of dosimetry protocols and standards for radiobiological studies is self-evident. Several protocols have been proposed for dose ...

Construction of a Realistic, Whole-Body, Three-Dimensional Equine Skeletal Model using Computed Tomography Data

Therapies based upon whole-body biomechanical assessments are successful for injury prevention and rehabilitation in human athletes. Similar approaches have rarely been used to study equine athletic injury. Degenerative osteoarthritis caused by mechanical stress can originate from chronic postural dysfunction, which, because the primary dysfunction is often distant from the site of tissue injury, is best identified through modeling whole-body biomechanics. To characterize whole-body equine kinematics, a realistic skeletal model of a horse was created from equine computed tomography (CT) data that can be used for functional anatomical and biomechanical modeling. Equine CT data were reconstructed into individual three-dimensional (3D) data sets (i.e., bones) using 3D visualization software and assembled into a complete 3D skeletal model. The model was then rigged and animated using 3D animation and modeling software. The resulting 3D skeletal model can be used to characterize equine postures associated with degenerative tissue changes as well as to identify postures that reduce mechanical stress at the sites of tissue injury. In addition, when animated into 4D, the model can be used to demonstrate unhealthy and healthy skeletal movements and can be used to develop preventative and rehabilitative individualized therapies for horses with degenerative lamenesses. Although the model will soon be available for download, it is currently in a format that requires access to the 3D animation and modeling software, which has quite a learning curve for new users. This protocol will guide users in (1) developing such a model for any organism of interest and (2) using this specific equine model for their own research questions.

The purpose of this protocol is to describe the method of creation of a realistic, whole-body, skeletal model of a horse that can be used for functional anatomical and biomechanical modeling to characterize whole-body mechanics.

使用计算机断层扫描数据构建逼真的全身三维马骨骼模型

Therapies based upon whole-body biomechanical assessments are successful for injury prevention and rehabilitation in human athletes. Similar approaches ...

Nanoparticle Tracking Analysis for the Quantification and Size Determination of Extracellular Vesicles

The physiological and pathophysiological roles of extracellular vesicles (EVs) have become increasingly recognized, making the EV field a quickly evolving area of research. There are many different methods for EV isolation, each with distinct advantages and disadvantages that affect the downstream yield and purity of EVs. Thus, characterizing the EV prep isolated from a given source by a chosen method is important for interpretation of downstream results and comparison of results across laboratories. Various methods exist for determining the size and quantity of EVs, which can be altered by disease states or in response to external conditions. Nanoparticle tracking analysis (NTA) is one of the prominent technologies used for high-throughput analysis of individual EVs. Here, we present a detailed protocol for quantification and size determination of EVs isolated from mouse perigonadal adipose tissue and human plasma using a breakthrough technology for NTA representing major advances in the field. The results demonstrate that this method can deliver reproducible and valid total particle concentration and size distribution data for EVs isolated from different sources using different methods, as confirmed by transmission electron microscopy. The adaptation of this instrument for NTA will address the need for standardization in NTA methods to increase rigor and reproducibility in EV research.

We demonstrate how to use a novel nanoparticle tracking analysis instrument to estimate the size distribution and total particle concentration of extracellular vesicles isolated from mouse perigonadal adipose tissue and human plasma.

细胞外囊泡的量化和尺寸测定纳米粒子跟踪分析

The physiological and pathophysiological roles of extracellular vesicles (EVs) have become increasingly recognized, making the EV field a quickly evolving ...

纳米草本涂层织物的抗菌功效

An Antimicrobial Fabric Using Nano-Herbal Encapsulation of Essential Oils

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and burns. These cotton-based protective coats provide ideal conditions for microbial growth and attachment sites due to their porous nature, moisture-holding capacity, and retention of warmth from the user&#39;s body. Several studies have demonstrated the survival of pathogenic bacteria on hospital garments and lab coats, acting as vectors of microbial transmission.
A common approach to fix these problems is the application of antimicrobial agents in textile finishing, but concerns have been raised due to the toxicity and environmental effects of many synthetic chemicals. The ongoing pandemic has also opened a window for the investigation of effective antimicrobials and eco-friendly and toxic-free formulations. This study uses two natural bioactive compounds, carvacrol and thymol, encapsulated in chitosan nanoparticles, which guarantee effective protection against four human pathogens with up to a 4-log reduction (99.99%). These pathogens are frequently detected in lab coats used in biohazard laboratories.
The treated fabrics also resisted up to 10 wash cycles with 90% microbial reduction, which is sufficient for the intended use. We made modifications to the existing standard fabric tests to better represent the typical scenarios of lab coat usage. These refinements allow for a more accurate evaluation of the effectiveness of antimicrobial lab coats and for the simulation of the fate of any accidental microbial spills that must be neutralized within a short time. Further studies are recommended to investigate the accumulation of pathogens over time on antimicrobial lab coats compared to regular protective coats.

作者聚焦：使用精油纳米草药封装的抗菌织物  

Antimicrobial lab coats prevent the cross-contamination of pathogen accumulation and accidental bio-spills. Here, we describe the protocol for developing a skin-friendly antimicrobial fabric using nano-herbal encapsulation and modified standard tests to precisely evaluate the efficacy and suitability for typical usage of the lab coat.

一种使用纳米草本封装精油的抗菌织物

Lab coats are widely used in biohazard laboratories and healthcare facilities as protective garments to prevent direct exposure to pathogens, spills, and ...