Name: evaluating dryocosmus kuriphilus-induced damage on castanea sativa
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作者感谢小行政区提契诺的森林服务和联邦政府机关为环境 FOEN 为这项研究部份地资助。

1. 树的选择和评估设计
<ol><li>如有可能, 通过使用可靠来源确定栗瘿栗瘿和Torymus的到来年份, 确定研究区的流行阶段 ( 例如,科学出版物、森林服务、板栗林管理人员的知识)。<ol>
<li>如果没有可靠的来源, 通过计算中华民国寄生率和下文所述的实地观察, 确定四主要流行阶段 (早期、峰值、恢复、生物防治)。<ol>
<li>鉴定阶段为早期流行时, 树冠显示既无显著损害, 也无冠透明, 当年瘿罕见,中华寄生是非常低或微不足道。</li>
			<li>当树冠显示高度透明度时, 确定流行峰, 虽然死枝稀少, 当年瘿丰富。</li>
			<li>在当前和往年的瘿丰富 (三年前) 和T. 中华寄生仍然很低或没有的情况下, 确定长期流行高峰。树冠仍然显示高透明度和另外的损伤由死的分支的第一个证据代表在冠。</li>
			<li>确定中华民国寄生率大于 75%10的早期恢复期。损伤仍然很大, 但当年瘿的数量减少, 一些分支产生无胆芽, 特别是从顶端芽。</li>
			<li>确定恢复阶段当中华民国寄生率永久大于 75%, 当年瘿是罕见的, 通常只限于单株树木, 大多数分支产生无胆芽。过去几年的瘿在老分支和死的分支由于D. 栗瘿攻击仍然是可看见的。</li>
			<li>确定完全恢复阶段时, 损坏和瘿 (过去和现在的年份) 是罕见的或缺席, 冠完全恢复。在严重受损的树木中, 前栗瘿攻击的一些痕迹 (例如,死芽或腐烂的过去几年的瘿) 仍可存在于冠内。 注: 补充文件1显示每个流行阶段的模范树冠图片。</li>
		</ol>
</li>
	</ol>
</li>
	<li>观察整个地区的板栗树, 以直观地估计树木之间和内部的损伤变异性。损伤变异通常是低的在早期的流行病和恢复阶段 (冠基本地是健康的) 并且在流行性高峰期间 (全冠是充分的瘿)。相比之下, 在中间流行阶段, 变异往往很高, 因为过去D. 栗瘿攻击的死芽仍然存在。</li>
	<li>根据1.1 和 1.2, 确定要分析的树数。不幸的是, 对于样本大小的具体规则是不可能的, 也不合适, 因为这可能因实地和/或研究目标的具体流行情况而异。根据我们10年的经验, 对于一个10公顷的网站, 我们建议如下 (也见表 1了解详细信息):<ol>
<li>每个站点至少取样十棵树, 而不考虑疫情阶段。虽然在早期流行病和恢复阶段三棵树是足够的, 增加样本大小到十将提供更多的统计能力的结果。</li>
		<li>在早期流行和恢复阶段, 每棵树取样一个分支。</li>
		<li>在疫峰期间, 如果瘿在树冠内均匀分布, 或每棵树上有两个树枝, 则每棵树取样一根树枝, 如果你注意到一些冠部分受到更严重的攻击。</li>
		<li>在其他流行阶段, 根据每棵树的树冠损伤的变异性, 将分支的数量增加到两个 (用于恢复良好的树木) 或三 (用于更多损坏的树木)。</li>
	</ol>
</li>
</ol>2. 外地的数据收集
<ol><li>准备适当的设备, 包括剪贴板, 露营椅, 整枝剪刀一般, 伸缩树剪, 30 米测量磁带, 和爬树设备, 如果顶冠超过8米需要分析。</li>
	<li>选择最具代表性的分支, 试图按比例覆盖树枝多样性在树冠 (健康的树木通常有类似的分支, 而受损的树木可能有不同程度的损害分支)。例如, 如果您选择收集每棵树三个分支, 收集最损坏的分支, 最健康和中间一个。</li>
	<li>只要可能, 选择仅建筑分支, 同时避免 reiterations (树干吸盘或 reiterations Hallé)16。</li>
	<li>确保分支机构长度至少为50厘米, 至少有10枝。</li>
	<li>在伸缩树剪的刀片附近附上测量磁带的开始, 以测量树枝在切削点上的高度。</li>
	<li>用伸缩剪切割分支, 记录其切割高度、侧面、类型 (结构或重复), 并用整枝剪刀一般细化分支选择, 以仅保留部件进行分析。</li>
	<li>为分支分配唯一 ID, 并记录其年龄、其最大长度 (从第一个分支点到顶点) 以获得一般信息。</li>
	<li>快速查看整个分支, 以获得其历史和当前状态的第一印象 (严重攻击与否), 并在图 2和图 3的帮助下确定所有重要的元素和功能, 以计算 DCI。</li>
</ol>3. 分支特征定义
以下定义是部分或完全复制从 Gehring等。20185, 以 Springer-出版社柏林海得尔堡2017的允许。
<ol><li>将芽(在拍摄上) 定义为一种新形成的芽, 在当前的营养季节从一枝成熟的芽上生长。</li>
	<li>根据取样日期 (例如,取样季节 = 2017, 在2016年生长的芽) 定义一个芽作为萌芽从早先植物季节。可以是死是活。</li>
	<li>将死射(Sd) 定义为栗瘿攻击后或因自然死亡而死亡。</li>
	<li>定义一个活的拍摄(as) 作为一个活生生的拍摄, 不要混淆与重新激活休眠芽。</li>
	<li>将重新激活的休眠芽(Bdor) 定义为一种新形成的芽, 在当前的营养季节从休眠的芽上生长到比拍摄年龄长的分支部分。</li>
	<li>在芽 (Gons) 上定义一种瘿, 如在基部或沿萌芽轴上发育的瘿。从技术上讲, 这些应该被称为 "瘿", 但为了与现存的文学保持一致, 我们将它们称作 "瘿"。 注:图 2和图 3显示了所选分支特征的示例。更详细和完整的描述 (这是超出本文的范围) 可能会发现在 Gehring等。20185和 Maltoni等。20124。</li>
</ol>4. 分支分析
<ol><li>计数和记录所有的活芽 (活芽)。</li>
	<li>计数并记录所有的死芽。</li>
	<li>计数并记录所有重新激活休眠芽。</li>
	<li>计数和记录所有的瘿在拍摄。 注意: 补充文件2显示了字段取样表单的示例, 补充文件3显示了填写的取样表单。</li>
</ol>5. 损伤综合指数的计算
<ol><li>计算死芽的比例 (Sd) 从死亡芽的数量除以芽的总数量 (死芽 + 活芽)。</li>
	<li>计算重新激活休眠芽 (BdoR) 的比例, 从重新激活休眠芽的数量除以活芽的总数 (BdoR + 活芽)。</li>
	<li>计算瘿的平均数量 (Gons) 从瘿的数量除以活芽的数量 (BdoR + 活芽)。</li>
	<li>使用公式计算 dci = (Sd * 0.479 + BdoR * 0.525 + Gons * 0.120) * 100。</li>
	<li>使用表 2评估损坏严重性。 注: 具有 DCI 功能的 R 脚本可在补充编码文件1中使用。它的描述在补充文件4中找到。</li>
</ol>

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作者没有什么可透露的。

栗瘿栗瘿在板栗树芽上产卵, 诱导春季瘿的形成。重复和失控的栗瘿攻击导致, 除了胆的形成, 一般分支腐败, 包括许多芽死亡和绿色光合叶面积的显著损失5。树木通常通过尝试通过激活休眠芽来产生替代芽来做出反应。因此, 特别是在疫情高峰期和恢复阶段, 经典女佣 (仅基于瘿丰度) 往往低估了栗瘿所造成的实际损害, 而 DCI 不仅基于瘿丰, 而且还死于芽和活?...

板栗 gallwasp栗瘿栗瘿Yasumatsu (膜翅目: Cynipidae) 是锥1、2、3属最显著的全球害虫。它通过反复的擦伤活动, 防止和抑制正常的芽发育4,5, 导致叶面积逐渐减少, 并由此造成树木绿色生物量和活力5,6 的损失., 休眠芽重新激活5和 gallwasp 后出现分支死亡率7,8增加。
欧洲 gallwasp 流行病的经验表明, 不受控制的和重复的 gallwasp 攻击可能会导致板栗 (锥磨) 的严重的冠层腐败。这可能导致冠叶面积的损失高达 70%, 既不是由激活休眠芽产生的替代叶补偿, 也不通过建立在同一植被期间的第二次冲洗5。
通过天然拮抗剂寄生蜂Torymus 中华Kamijo (膜翅目: Torymidae)9、10, 唯一成功的方法是减少害虫种群, 使板栗树恢复正常。一旦生物控制通过其天敌实现, 板栗树开始产生新的健康芽。如果树损伤水平是非常高的, 这可能从终端芽开始仅发生, 由于事实它通常无侵扰由于它的形成在 gallwasp 产卵活动4以后。这意味着一个长的恢复过程之前, 整个树冠重新建立5。
为了检验板栗树在Torymus生物防治后的阳性反应, 并验证 sylvicultural (修剪、细化) 干预的必要性, 森林管理人员和板栗种植者需要一种快速、通过对害虫的拮抗剂进行生物控制后, 从最初的 gallwasp 流行阶段, 对损伤程度及相关分支体系结构和叶面积演化进行可靠的评估。迄今已开发并使用了几种评估 gallwasp 感染程度 (女仆) 的方法, 例如测量被攻击芽11的比例或每芽12的平均瘿数。女佣不直接测量绿色生物量 (如叶面积), 后备结构, 如休眠芽, 反应结构 (例如,重新激活休眠芽和第二冲洗), 或前一年的损害 (如死芽) 作为主要当前树生命力和活力的代理6,13,14。此外, 大多数女佣只是根据在树枝上发现的瘿的数量和低估了真正的分支损害, 特别是在害虫流行的高峰期 (图 1)。
本文描述了 Gehring 的损伤复合指数 (DCI) 方法. 20185认为绿色生物量的代理, 储备如休眠芽和树反应 (休眠芽恢复和第二冲洗), 使一个现实的, 可靠的, 并合理快速评估的损害, 通过所有阶段的流行病,特别是结合 Gehring 提出的评估努力优化. 201715。
具体而言, 本文的目标是 1), 详细说明外地议定书, 包括将要评估的相关分支特征, 2) 提出损害综合指数公式和 3, 提出改进的严重性等级换算为 DCI。

<table><tbody><tr><td>Name of Material/ Equipment</td><td>Company</td><td>Catalog Number</td><td>Comments</td></tr><tr><td>Clipboard</td><td>Any brand</td><td></td><td></td></tr><tr><td>Camping chair&lt;br/&gt; (Foldable and lightweight chair)</td><td>Any brand</td><td></td><td>Companies: Kelty, Campz, Half-Ton.</td></tr><tr><td>Felco 9 secateurs&lt;br/&gt; (One-hand pruning shear)</td><td>Felco</td><td></td><td>Other companies: Bahco.</td></tr><tr><td>AP-5M-Aluminium Pole&lt;br/&gt; (Telescopic tree pruner pole)</td><td>Bahco</td><td>8152079</td><td>Other companies: Spear &amp;amp; Jackson, Kingfisher, Hortex, Fiskars.</td></tr><tr><td>P34-37 top pruner&lt;br/&gt; (Telescopic tree pruner head)</td><td>Bahco</td><td>8002787</td><td></td></tr><tr><td>100 ft Fiberglass Long Tape&lt;br/&gt; (30 m measuring tape)</td><td>Stanley</td><td>34-790</td><td>Other companies: Tjima, Freemans, Astor, Lux.</td></tr><tr><td>Parallel 10.5mm&lt;br/&gt; (Low stretch kernmantel rope, flexible and lightweight for rope access)</td><td>Petzl</td><td>R077AA03</td><td>Basic equipment for tree climbing&amp;nbsp; (if necessary). Many other equipment configurations can be used for tree climbing, depending on the situation and on single operator preferences. We used Pezl equipment but many other companies offer similar products (e.g. Edelrid, Notch, Climbing technologies, DMM, ...). For a complete overview of equipment and companies we recommend a search in google&amp;nbsp; &quot;tree climbing gear&quot; as search keyword. PLEASE NOTE: tree climbing activities should be done only by professionals and are submitted to specific regulatory prescriptions according to the country.</td></tr><tr><td>Avao Sit&lt;br/&gt; (Harness for work positioning and suspension)</td><td>Petzl</td><td>C69AFA 2</td><td></td></tr><tr><td>Rig&lt;br/&gt; (Compact self-braking descender)</td><td>Petzl</td><td>D21A</td><td></td></tr><tr><td>Ascension&lt;br/&gt; (Handled rope clamp for rope ascents)</td><td>Petzl</td><td>B17ALA</td><td></td></tr><tr><td>Eclipse&lt;br/&gt; (Storage for throw-line)</td><td>Petzl</td><td>S03Y</td><td></td></tr><tr><td>Airline&lt;br/&gt; (Throw-line)</td><td>Petzl</td><td>R02Y 060</td><td></td></tr><tr><td>Jet&lt;br/&gt; (Throw-bag)</td><td>Petzl</td><td>S02Y 300</td><td></td></tr><tr><td>Vertex best&lt;br/&gt; (Comfortable helmet for work at height and rescue)</td><td>Petzl</td><td>A10BYA</td><td></td></tr><tr><td>Zillon&lt;br/&gt; (Adjustable work positioning lanyard for tree care)</td><td>Petzl</td><td>L22A 040</td><td></td></tr><tr><td>Ok&lt;br/&gt; (Lightweight oval carabiner)</td><td>Petzl</td><td>M33A SL</td><td></td></tr></tbody></table>

evaluating dryocosmus kuriphilus-induced damage on castanea sativa

栗瘿栗瘿Yasumatsu 已成为Castaneasativa的主要害虫, 因为它的到来在欧洲。其擦伤活性导致不同类型的胆类形成, 防止正常芽的发育。重复和失控的攻击造成, 除了生产瘿和随之而来的与胆汁相关的减少叶面积, 逐步腐败的分支体系结构, 包括死亡的分支部分, 并增加休眠芽激活。迄今为止, 很少有人试图量化分支体系结构的损坏。此外, 评估已发展的侵害程度 (女仆) 的不同方法只注重瘿的存在和富足。
利用叶面积与边材区域的关系作为绿色生物量指标, 我们在以前的研究中开发了一种损伤复合指数 (DCI), 它考虑了最重要的分支结构特征, 允许在整个流行病过程。
本研究的目的是提出这一新的方法, 并强调不同的损害描述与其他广泛使用的指数。结果显示, 与女仆相比, 协会对分支损伤的描述是更好的, 尤其是在流行高峰时期。最后, 我们建议如何通过我们的综合损伤指数、使用经典方法的侵扰程度以及冠层透明度评估来正确评估害虫的总体影响。

共计25个现场在提契诺, 瑞士被参观了在2013和2016之间为了创造一个世俗梯度覆盖所有瘿蜂流行性阶段。总计, 我们收集和分析了94个分支在5个站点在早期的流行性阶段 (害虫的到来和树损伤的开始), 200 个分支在5个站点在流行性高峰 (中等到严重损伤由于高水平的D. 栗瘿攻击), 200 个分支在5个站点在恢复阶段 (由中华民国的生物防治到达和开始渐进的树恢复), 并?...

Watch this Scientific Journal Video about Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa at JoVE.com

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

栗瘿栗瘿致锥害的评价

通过考虑瘿的丰度, 而不是考虑到相关的分支腐败, 来评估栗瘿栗瘿造成的损害是普遍的做法。我们提出一个综合损伤指数, 考虑到最重要的分支特征, 从而使更现实的损害评估。

Dryocosmus kuriphilus Yasumatsu has become a major pest for Castaneasativa since its arrival in Europe. Its galling activity results in the formation of ...

evaluating-dryocosmus-kuriphilus-induced-damage-on-castanea-sativa

Research

JoVE Journal

Environment

Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

7.2K 次观看.  WSL Swiss Federal Research Institute. 通过考虑瘿的丰度, 而不是考虑到相关的分支腐败, 来评估栗瘿栗瘿造成的损害是普遍的做法。我们提出一个综合损伤指数, 考虑到最重要的分支特征, 从而使更现实的损害评估。

视频: Evaluating Dryocosmus Kuriphilus-induced Damage on Castanea Sativa

Optimization of the Wound Scratch Assay to Detect Changes in Murine Mesenchymal Stromal Cell Migration After Damage by Soluble Cigarette Smoke Extract

Cell migration is vital to many physiological and pathological processes including tissue development, repair, and regeneration, cancer metastasis, and inflammatory responses. Given the current interest in the role of mesenchymal stromal cells in mediating tissue repair, we are interested in quantifying the migratory capacity of these cells, and understanding how migratory capacity may be altered after damage. Optimization of a rigorously quantitative migration assay that is both easy to customize and cost-effective to perform is key to answering questions concerning alterations in cell migration in response to various stimuli. Current methods for quantifying cell migration, including scratch assays, trans-well migration assays (Boyden chambers), micropillar arrays, and cell exclusion zone assays, possess a range of limitations in reproducibility, customizability, quantification, and cost-effectiveness. Despite its prominent use, the scratch assay is confounded by issues with reproducibility related to damage of the cell microenvironment, impediments to cell migration, influence of neighboring senescent cells, and cell proliferation, as well as lack of rigorous quantification. The optimized scratch assay described here demonstrates robust outcomes, quantifiable and image-based analysis capabilities, cost-effectiveness, and adaptability to other applications.

The capacity to migrate is a key function of many different cell types, including mesenchymal stromal cells (MSCs). However, quantifying alterations in migratory capacity after damage is challenging. This protocol describes an easily adaptable migration assay that uses rigorous statistics to quantify changes in MSC migratory capacity after damage.

伤口划痕试验的优化来探测在小鼠间充质干细胞迁移的可溶性香烟烟雾提取物受损后

Cell migration is vital to many physiological and pathological processes including tissue development, repair, and regeneration, cancer metastasis, and ...

科研

发育生物学

Remote Sensing Evaluation of Two-spotted Spider Mite Damage on Greenhouse Cotton

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the two-spotted spider mite (TSSM), Tetranychus urticae Koch, on greenhouse grown cotton. TSSM is a polyphagous pest which occurs on a variety of field and horticultural crops. It often becomes an early season pest of cotton in damaging proportions as opposed to being a late season innocuous pest in the mid-southern United States. Evaluation of acaricides is important for maintaining the efficacy of and preventing resistance to the currently available arsenal of chemicals and newly developed control agents. Enumeration of spider mites for efficacy evaluations is laborious and time consuming. Therefore, subjective visual damage rating is commonly used to assess density of spider mites. The NDVI (Normalized Difference Vegetation Index) is the most widely used statistic to describe the spectral reflectance characteristics of vegetation canopy to assess plant stress and health consequent to spider mite infestations. Results demonstrated that a multispectral optical sensor is an effective tool in distinguishing varying levels of infestation caused by T. urticae on early season cotton. This remote sensing technique may be used in lieu of a visual rating to evaluate insecticide treatments.

This manuscript describes a multispectral optical sensor that effectively detected damage to early season cotton artificially infested with varying densities of the two-spotted spider mite populations.

温室棉二斑叶螨危害的遥感评价

The objective of this study was to evaluate a ground-based multispectral optical sensor as a remote sensing tool to assess foliar damage caused by the ...

环境科学

Impact Assessment of Repeated Exposure of Organotypic 3D Bronchial and Nasal Tissue Culture Models to Whole Cigarette Smoke

Cigarette smoke (CS) has a major impact on lung biology and may result in the development of lung diseases such as chronic obstructive pulmonary disease or lung cancer. To understand the underlying mechanisms of disease development, it would be important to examine the impact of CS exposure directly on lung tissues. However, this approach is difficult to implement in epidemiological studies because lung tissue sampling is complex and invasive. Alternatively, tissue culture models can facilitate the assessment of exposure impacts on the lung tissue. Submerged 2D cell cultures, such as normal human bronchial epithelial (NHBE) cell cultures, have traditionally been used for this purpose. However, they cannot be exposed directly to smoke in a similar manner to the in vivo exposure situation. Recently developed 3D tissue culture models better reflect the in vivo situation because they can be cultured at the air-liquid interface (ALI). Their basal sides are immersed in the culture medium; whereas, their apical sides are exposed to air. Moreover, organotypic tissue cultures that contain different type of cells, better represent the physiology of the tissue in vivo. In this work, the utilization of an in vitro exposure system to expose human organotypic bronchial and nasal tissue models to mainstream CS is demonstrated. Ciliary beating frequency and the activity of cytochrome P450s (CYP) 1A1/1B1 were measured to assess functional impacts of CS on the tissues. Furthermore, to examine CS-induced alterations at the molecular level, gene expression profiles were generated from the tissues following exposure. A slight increase in CYP1A1/1B1 activity was observed in CS-exposed tissues compared with air-exposed tissues. A network-and transcriptomics-based systems biology approach was sufficiently robust to demonstrate CS-induced alterations of xenobiotic metabolism that were similar to those observed in the bronchial and nasal epithelial cells obtained from smokers.

The aim of this protocol is to expose human organotypic 3D bronchial and nasal tissue models to mainstream cigarette smoke (CS) at the air-liquid interface. The impact of CS on the tissues is then investigated using a cytochrome P450 activity assay, a cilia beating measurement, and a systems biology approach.

反复接触器官的3D支气管及鼻部组织文化模型对整个卷烟烟雾的影响评估

Cigarette smoke (CS) has a major impact on lung biology and may result in the development of lung diseases such as chronic obstructive pulmonary disease ...

生物工程

Ecotoxicological Methodologies to Evaluate Biomarkers at Different Scales in Neotropical Anurans

The new questions in ecotoxicology highlight the importance of applying a battery of biomarkers, as this results in ecotoxicological predictions that improve not only the interpretation of the effects of environmental stressors on organisms but also the determination of their possible impact. It is well known that the use of ecotoxicological biomarkers at different levels of organization allows for the prediction of the biological responses of organisms to environmental stressors, which is useful in environmental risk assessment. 
Nevertheless, it is necessary to consider the optimization of basic procedures, to generate historical data in control groups, and to employ specific bioassays to evaluate responses in organs and tissues in order to elucidate the nature and variation of the effects observed. Therefore, the present work aims to describe several ecotoxicological methodologies employed in all stages of neotropical anurans at different ecological levels and to validate them as useful biomarkers to be used both in wildlife and in laboratory conditions. In this work, these biomarkers were applied at the individual/organismic level (body condition index), histological/physiological level (histopathology, histometric, and pigmentary analyses), biochemical level (oxidative stress enzymes), and genetic level (direct and oxidative damage in DNA by comet assay). 
Although these methodologies have small variations or modifications depending on the species, these techniques provide effective biomarkers for evaluating the effect of xenobiotics on anurans, which possess certain characteristics that make them useful indicator species of aquatic and terrestrial ecosystems. In conclusion, the battery of biomarkers employed in the present study has proven to be adequate for estimating toxic responses in Neotropical anurans and can be further recommended as bioindicators for identifying the impact of pollutants on the aquatic ecosystems of the region. Finally, it is recommended to achieve the standardization of these important biomarkers for anurans in specific regions as well as to possibly include them in risk assessments and decision-making.

<meta charset="utf8"></head><body>评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

In this paper, standardized ecotoxicological methods for the evaluation of biomarkers in neotropical anuran species are presented. Specifically, this paper details several methodologies at different scales of ecotoxicological evaluation, such as the genetic, cellular-histological, biochemical, morphological, and individual levels.

评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

The new questions in ecotoxicology highlight the importance of applying a battery of biomarkers, as this results in ecotoxicological predictions that ...

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. Radioactive beams can penetrate through the skin and affect the structures in their path including skin, muscles, and internal organs. Skin is the first structure to get exposed to radiation and is susceptible to develop chronic fibrosis, which is challenging to treat. Currently, limited treatment options show moderate efficacy in mitigating radiation-related skin fibrosis. A key factor hindering the development of effective countermeasures is the absence of a convenient and robust model that could allow for translation of the experimental findings to humans. Here, a robust and reproducible murine hind limb skin fibrosis model has been established for prophylactic and therapeutic evaluation of possible agents for functional and molecular recovery.
The right hind limb was irradiated using a single dose of 40 (Gray) Gy to induce skin fibrosis. Subjects developed edema and dermatitis in the early stages proceeded by visible skin constriction. Irradiated limbs showed a significantly reduced limb range of motion in the following weeks. In late stages, acute side effects subsided, yet chronic fibrosis persisted. A gait index was performed as an additional functional assay, which demonstrated the development of functional impairment. These non-invasive methods demonstrated reliable measurements for tracing fibrosis progression, which is supported by histological analyses. The radiation dose, application, and post-irradiation analyses employed in this model offer a vigorous and reproducible method for studying radiation-induced skin fibrosis and testing the efficacy of therapeutical agents.

Here we present a protocol to induce radiation-induced skin fibrosis in the hind limb of mice and perform post-irradiation measurements of chronic impairment via limb excursion and gait index analyses to evaluate the functional outcome. The model elucidates radiation-related skin fibrosis mechanisms and is useful in subclinical therapeutic studies.

Radiation-induced skin fibrosis (RISF) can result from a plethora of scenarios including cancer therapy, accidental exposure, or acts of terrorism. ...

癌症研究

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain perception in rodents. Withdrawal thresholds can be measured first at baseline and then following drug, venom, injury, allergen,&#160;or otherwise evoked inflammation by applying an accurate force on very specific areas of the skin. An electronic von Frey apparatus allows precise assessment of mouse hind paw withdrawal thresholds that are not limited by the available filament sizes in contrast to classical von Frey measurements. The ease and rapidity of measurements allow for incorporation of assessment of tactile sensitivity outcomes in diverse models of rapid-onset inflammatory and neuropathic pain as multiple measurements can be taken within a short time period. Experimental measurements for individual rodent subjects can be internally controlled against individual baseline responses and exclusion criteria easily established to standardize baseline responses within and across experimental groups. Thus, measurements using an electronic von Frey apparatus represent a useful modification of the well-established classical von Frey filament-based assays for rodent mechanical allodynia that may also be applied to other nonhuman mammalian models.

Electronic von Frey measurements are an effective and improved alternative to classical von Frey filaments, as they allow rapid and precise measurement of changes in rodent mechanical withdrawal thresholds to continuously applied pressure stimuli before and after an inflammatory event.

Measuring inflammation-induced changes in thresholds of hind paw withdrawal from mechanical pressure is a useful technique to assess changes in pain ...

神经科学

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila larvae, customized metal nickel-titanium alloy (nitinol) filaments. These mechanical probes are similar to the von Frey filaments used in vertebrates to measure mechanical nociception. Here, we demonstrate how to make and calibrate these mechanical probes and how to generate a full behavioral dose-response from subthreshold (innocuous or non-noxious range) to suprathreshold (low to high noxious range) stimuli. To demonstrate the utility of the probes, we investigated tissue damage-induced hypersensitivity in Drosophila larvae. Mechanical allodynia (hypersensitivity to a normally innocuous mechanical stimulus) and hyperalgesia (exaggerated responsiveness to a noxious mechanical stimulus) have not yet been established in Drosophila larvae. Using mechanical probes that are normally innocuous or probes that typically elicit an aversive behavior, we found that Drosophila larvae develop mechanical hypersensitization (both allodynia and hyperalgesia) after tissue damage. Thus, the mechanical probes and assay that we illustrate here will likely be important tools to dissect the fundamental molecular/genetic mechanisms of mechanical hypersensitivity.

An Improved Assay and Tools for Measuring Mechanical Nociception in Drosophila Larvae

The goal of this protocol is to show how to perform an improved assay for mechanical nociception in Drosophila larvae. We use the assay here to demonstrate that mechanical hypersensitivity (allodynia and hyperalgesia) exists in Drosophila larvae.

测量 果蝇 拉瓦的机械结节的改进的检测和工具

Published assays for mechanical nociception in Drosophila have led to variable assessments of behavior. Here, we fabricated, for use with Drosophila ...

行为学

Electroantennographic Bioassay as a Screening Tool for Host Plant Volatiles

Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host plant.1,2 When a host plant is an important agronomic commodity feeding damage by insect pests can inflict serious economic losses to growers. Accordingly, kairomones can be used as attractants to lure or confuse these insects and, thus, offer an environmentally friendly alternative to pesticides for insect control.3 Unfortunately, plants can emit a vast number volatiles with varying compositions and ratios of emissions dependent upon the phenology of the commodity or the time of day. This makes identification of biologically active components or blends of volatile components an arduous process. To help identify the bioactive components of host plant volatile emissions we employ the laboratory-based screening bioassay electroantennography (EAG). EAG is an effective tool to evaluate and record electrophysiologically the olfactory responses of an insect via their antennal receptors. The EAG screening process can help reduce the number of volatiles tested to identify promising bioactive components. However, EAG bioassays only provide information about activation of receptors. It does not provide information about the type of insect behavior the compound elicits; which could be as an attractant, repellent or other type of behavioral response. Volatiles eliciting a significant response by EAG, relative to an appropriate positive control, are typically taken on to further testing of behavioral responses of the insect pest. The experimental design presented will detail the methodology employed to screen almond-based host plant volatiles4,5 by measurement of the electrophysiological antennal responses of an adult insect pest navel orangeworm (Amyelois transitella) to single components and simple blends of components via EAG bioassay. The method utilizes two excised antennae placed across a "fork" electrode holder. The protocol demonstrated here presents a rapid, high-throughput standardized method for screening volatiles. Each volatile is at a set, constant amount as to standardize the stimulus level and thus allow antennal responses to be indicative of the relative chemoreceptivity. The negative control helps eliminate the electrophysiological response to both residual solvent and mechanical force of the puff. The positive control (in this instance acetophenone) is a single compound that has elicited a consistent response from male and female navel orangeworm (NOW) moth. An additional semiochemical standard that provides consistent response and is used for bioassay studies with the male NOW moth is (Z,Z)-11,13-hexdecadienal, an aldehyde component from the female-produced sex pheromone.6-8

A method to rapidly screen host plant volatiles by measurement of the electrophysiological response of adult navel orangeworm (Amyelois transitella) antennae to single components and blends via electroantennographic analysis is demonstrated.

作为寄主植物挥发物的筛选工具electroantennographic生物活性

Plant volatiles play an important role in plant-insect interactions. Herbivorous insects use plant volatiles, known as kairomones, to locate their host ...

生物学

Systematic Scoring Analysis for Intestinal Inflammation in a Murine Dextran Sodium Sulfate-Induced Colitis Model

Murine colitis models are tools that are extensively employed in studies focused on understanding the pathobiology of inflammatory intestinal disorders. However, robust standards for objective and reproducible quantification of disease severity remain to be defined. Most colitis analysis methods rely on limited histological scoring of small segments of intestine, leading to partial or biased analyses. Here, we combine high-resolution image acquisition and longitudinal analysis of the entire colon to quantify intestinal injury and ulceration in the dextran sodium sulfate (DSS) induced model of murine colitis. This protocol allows for the generation of objective and reproducible results without extensive user training. Here, we provide comprehensive details on sample preparation and image analysis using examples of data from DSS induced colitis. This method can be easily adapted to other models of murine colitis that have significant inflammation associated with mucosal injury. We demonstrate that the fraction of inflamed/injured and eroded/ulcerated mucosa relative to the complete length of the colon closely parallels clinical findings such as weight loss amid DSS-induced disease progression. This histological protocol provides a reliable time and cost-effective aid to standardize analyses of disease activity in an unbiased way in DSS colitis experiments.

Systematic scoring of intestinal inflammation using a free computer-assisted system is a powerful tool to quantitatively compare histopathological changes in colitis models characterized by the presence of ulcers and inflammatory changes. Histological colitis score evaluation strengthens clinical observations and facilitates data interpretation.

穆林·德克斯特兰硫酸钠诱导结肠炎模型中肠道炎症的系统评分分析

Murine colitis models are tools that are extensively employed in studies focused on understanding the pathobiology of inflammatory intestinal disorders. ...

栗瘿栗瘿致锥害的评价

摘要

探索更多视频

此视频中的章节

伤口划痕试验的优化来探测在小鼠间充质干细胞迁移的可溶性香烟烟雾提取物受损后

温室棉二斑叶螨危害的遥感评价

反复接触器官的3D支气管及鼻部组织文化模型对整个卷烟烟雾的影响评估

评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

测量果蝇拉瓦的机械结节的改进的检测和工具

作为寄主植物挥发物的筛选工具electroantennographic生物活性

穆林·德克斯特兰硫酸钠诱导结肠炎模型中肠道炎症的系统评分分析

栗瘿栗瘿致锥害的评价

摘要

探索更多视频

此视频中的章节

伤口划痕试验的优化来探测在小鼠间充质干细胞迁移的可溶性香烟烟雾提取物受损后

温室棉二斑叶螨危害的遥感评价

反复接触器官的3D支气管及鼻部组织文化模型对整个卷烟烟雾的影响评估

评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

评估新热带 Anurans 不同尺度生物标志物的生态毒理学方法

Establishment of a Robust and Reproducible Model of Radiation-Induced Skin and Muscle Fibrosis

Measuring Changes in Tactile Sensitivity in the Hind Paw of Mice Using an Electronic von Frey Apparatus

测量 果蝇 拉瓦的机械结节的改进的检测和工具

作为寄主植物挥发物的筛选工具electroantennographic生物活性

穆林·德克斯特兰硫酸钠诱导结肠炎模型中肠道炎症的系统评分分析

测量果蝇拉瓦的机械结节的改进的检测和工具