Name: a rapidly incremented tethered-swimming maximal protocol for cardiorespiratory assessment of swimmers
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Description: Watch this Scientific Journal Video about A Rapidly Incremented Tethered-Swimming Maximal Protocol for Cardiorespiratory Assessment of Swimmers at JoVE.com

这项工作得到了葡萄牙CIPER-科学和技术基金会（CFCT）的支持（UID/DTP/00447/2019），部分资金来自巴西高级科学基金会，其中部分资金来自巴西国家科学与技术基金会（CAPES）的资助。 和圣保罗研究基金会 - FAPESP （PROCESS 2016/04544-3 和 2016/17735-1）。作者感谢若昂·吉列尔梅·德奥利维拉在数据采样方面给予的帮助。Mério A. C. Espada 感谢葡萄牙体育和青年学院 IPDJ 的财政支持。

从下述代表性主题数据中提取的研究中，参与者须在实验程序、相关风险和参与的潜在好处解释后，在开始测试之前给予书面知情同意。第一次访问包括一次熟悉会议，在培训期间，游泳者被介绍到系绳游泳的概念和在实际测试中有效的测量技术。在第二次访问期间进行了全方位的系绳游泳测试，并在第三次访问中执行了快速增加的系绳游泳协议。两项测试均在半奥林匹?...

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一般来说，从INC对耐力运动员的评估中得出的主要兴趣参数是V_O2max，它用于监测运动员的心肺健康水平。此外，V•O2max在规定训练计划时通常用作分配练习 WR 的方法（即，WR 指定为 V•O2max的百分比）。然而，越来越多的研究证实，肺气交换（以及，进而，肌肉代谢）对WR线性增加的反应不是线性的，重要的是，这种非线性的特征因不同的个体（以及...

当运动保持在恒定的工作速度（CWR）时，根据WR下降4，5的"运动强度域"，生理反应概况明显不同。具体来说，实现V_O2和血液[乳酸]"稳定状态"是快速在中度域，延迟在重域和无法实现在严重域4，5。众所周知，在INC（V[O2GET）期间，O2在GET的消耗率是CWR3、6期间将中度与重域分开的代谢率。虽然存在争议，但最近的一些观察表明，在RCP（V-O2RCP）中，O2的消耗速率与重/重分离7、8、9、10之间的等值相似。因此，从INC期间收集的数据中识别VO2GET和V+O2RCP，对于通过代谢率为耐力运动员开一个特定领域的训练方案可能很有用，并告诫说，根据增量测试8、11中产生的VO+2工作速率关系，将代谢率与特定工作速率对齐比简单地调整代谢率更为复杂。
当确定V&O2max的测试概念被初步探索时，研究人员让受试者在单独的第1天以越来越快的速度进行轨道跑步，达到运动耐受极限（Tlim）。研究随后证实，V*O2max也可以确定从类似的回合执行到Tlim在同一天与休息时间穿插12。最终，结果显示，具有WR的连续协议在特定时间间隔（例如，每3分钟）以增量方式增加，显示与不连续测试13相同的V+O2峰值。因此，这些"分级运动测试"成为确定心肺健康标准的标准。然而，在1981年，Whipp和他的同事发表了一项研究表明，为了V&O2max测量的目的，INC也可以完全在非稳定状态下进行;也就是说，随着WR的不断增加，作为一个"平滑的时间函数"（RAMP-INC）14 。与具有扩展阶段和每个阶段相对较大的 WR 增加的 INC 不同，在 RAMP-INC 期间逐渐增加可确保将 GET 和 RCP 分开的"等值缓冲区域"将明确定义15。此外，与具有分阶段的INC类似，RAMP-INC可用于评估"运动经济性"（即，每个给定WR所需的V•O2）;然而，与具有阶段的 INC 不同，在这种情况下， 用于此目的的"三角洲效率"（即 V[O2-WR 关系的斜率）的反面是11，考虑到由于 VO2对强度频谱工作速率的响应的复杂性，此参数本身不是 INC 的不可变特征（例如，从不同的基线工作速率启动的 RAMP-INC 或以不同斜坡斜率为特征）16.
对于一般体能测试，INC 通常在腿部测速仪或跑步机上进行，因为这些模式更可用，而腿部骑自行车和步行/跑步为普通人所熟悉。此外，管理RAMP-INC需要以较小的增量（例如，每2个s1W）持续增加WR的能力;因此，测速仪（通常为腿部循环）最适合此类测试。然而，运动员评估是更复杂的，因为运动员必须进行测试，同时执行其运动所需的特定运动模式。对于参加涉及跑步的运动的自行车运动员和个人来说，这没有问题，因为上述测试机器的可访问性和适用性。相反，在评估水生运动员时，通过气体交换和通风数据收集以及 RAMP-INC 所需的逐步 WR 递增进行生态有效的测试更具挑战性。
在自动收集系统出现之前，游泳者经常在进行最大游泳17次后使用道格拉斯袋收集进行气体交换评估。一旦自动化系统被开发出来，"实时"收集就发生了，但不是在"真实游泳"条件下（例如，游泳者在控制WR的烟道中游泳）17。不幸的是，前者由于"向后外推"的假设而具有固有的局限性，而后者则引起人们对泳泳改变技术17程度的担忧。目前的技术是使用便携式呼吸式收集系统，在自由泳17期间与游泳者在游泳池旁边移动。虽然这种类型的测量提高了生态有效性，但逐步WR递增具有挑战性。事实上，在自由游泳期间，INC 通常涉及以逐渐增加的速度14、15的设定距离间隔（例如 200 米）。这意味着测试由具有较大不相等的 WR 增量的长阶段组成。因此，使用这个测试18，19的研究人员报告只有一个代谢断点（通常称为"厌氧阈值"）也就不足为奇了。相反，我们最近已经表明，V[O2GET和 V-O2RCP都可以从收集的数据中确定，而游泳者在游泳池中进行静静游泳时，对逐渐和快速增加的负载（即增量系绳游泳）20。虽然游泳过程中存在的独特呼吸模式可能使上述断点与典型的评估模式（个人观察）相比更难识别，但我们认为，这种测试方法可能适合作为"游泳测速仪"，可用于对游泳者进行心肺评估，其方式类似于骑自行车者使用固定循环的方式。事实上，我们已经表明，V[O2GET，V[O2RCP和运动经济性（如V[O2-负载斜率所示）》都可以从20以下描述的快速递增的系绳游泳协议中确定。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>3-L syringe</td>
			<td>Hans Rudolph</td>
			<td></td>
			<td>Calibration device</td>
		</tr>
		<tr>
			<td>Aquatrainer</td>
			<td>COSMED</td>
			<td></td>
			<td>Snorkel system/gas-exchange measurement</td>
		</tr>
		<tr>
			<td>K4b2</td>
			<td>COSMED</td>
			<td></td>
			<td>Portable CPET unit/gas-exchange measurement</td>
		</tr>
		<tr>
			<td>N200PRO</td>
			<td>Cefise</td>
			<td></td>
			<td>Software program for analysis of force signal</td>
		</tr>
		<tr>
			<td>Pacer 2 Swim</td>
			<td>Kulzer TEC</td>
			<td></td>
			<td>Swimming velocity management/underwater LED line</td>
		</tr>
		<tr>
			<td>Tether-system</td>
			<td>Own design</td>
			<td></td>
			<td>Pulley-Rope system/loading management</td>
		</tr>
		<tr>
			<td>Tether attachment</td>
			<td>CEFISE</td>
			<td></td>
			<td>Bracket for attachment to swimmer</td>
		</tr>
	</tbody>
</table>

a rapidly incremented tethered-swimming maximal protocol for cardiorespiratory assessment of swimmers

增量运动测试是评估耐力运动员心肺能力的标准手段。虽然最大耗氧率通常用作这方面的标准测量，但从功能角度来看，两个代谢断点反映了乳酸盐生产/消耗动态的变化，因为工作速度的提高可能更适用于耐力运动员。运动经济性是衡量耐力运动员评价的一个重要参数，它代表了副极限工作表现的耗氧率。斜坡增量测试包括逐步但快速提高工作速度，直到达到运动容差的极限，对于确定这些参数非常有用。这种类型的测试通常在循环测速仪或跑步机上进行，因为在工作速率递增方面需要精度。但是，运动员在进行运动所需的锻炼模式时，应进行测试。因此，游泳者通常在自由泳增量测试中进行评估，而这种精度很难达到。我们最近建议，固定游泳对逐渐增加的负载（增量系绳游泳）可以作为一个"游泳测速计"，允许足够的精度，以适应一个渐进的，但快速的加载模式，揭示上述代谢断点和运动经济。然而，在这种协议期间达到的耗氧峰值速率与自由游泳期间测量的最大速率近似的程度仍有待确定。在本文中，我们将解释如何利用这种快速增加的系绳游泳方案来评估游泳者的心肺能力。具体来说，我们解释如何使用此协议对短距离竞技游泳者的评估显示，他的吸氧率分别为30.3 mL和34.8mL=min-1=kg-1 BM，在他的气体交换阈值和呼吸补偿点。

表1中所示和图1-4中所示的数据显示了男性游泳运动员（年龄，24岁）观察到的响应概况。在收集数据时，这位游泳运动员已经接受7年的竞技游泳训练。他的专长是短距离（即50米和100米自由泳）。
INC 上的初始负载设定在超过此游泳者在开始全能游泳 （F基） 之前保持身体对齐所需的负载，其?...

Watch this Scientific Journal Video about A Rapidly Incremented Tethered-Swimming Maximal Protocol for Cardiorespiratory Assessment of Swimmers at JoVE.com

A Rapidly Incremented Tethered-Swimming Maximal Protocol for Cardiorespiratory Assessment of Swimmers

与自由泳期间的测量（这会带来固有的挑战和局限性）不同，使用更可行、更易于管理系绳游泳的游泳者，可以确定游泳者心肺功能的重要参数。通过气体交换和通风数据收集快速增加的协议。

游泳运动员心肺评估的快速递增的绑带游泳最大协议

Incremental exercise testing is the standard means of assessing cardiorespiratory capacity of endurance athletes. While the maximal rate of oxygen ...

在静止游泳过程中，在不断增加的负载时，可以很容易辨别出气体交换阈值和呼吸补偿点，这是自由泳过程中以越来越快的速度测量的两个重要参数。增量运动期间通过增加负荷的强度增加可以是对称的、渐进的和快速的，在增量自由泳中，这种强度不是以越来越快的速度增加的。管理这个程序的将是莱安德罗·奥利维拉、路易斯·古斯塔沃·桑托斯和卡米拉·瓦斯康塞洛斯，这些研究生来自我的实验室。游泳运动员将是朱莉娅加藤谁已经参加了五年的竞技游泳训练。要准备 500 公斤的称重电池，用于测量游泳者在两次试验中可以施加的最高力，包括 30 秒的游泳，请打开 N2000PRO 软件程序，然后打开帮助菜单以验证计算机和称重细胞分析仪之间的通信链路。建立与 RS232 接口的连接后，将出现绿色信号。设置开始测试的倒计时、采样持续时间和休息间隔。然后将帧/秒设置为 100 赫兹。设置力度量单位，然后以毫秒为单位设置采集时间。然后，用游泳池外的游泳者校准负载为零和 10 公斤的称重。在开始两试测试之前，提供有关全前爬行游泳正确表现的说明，让游泳者在池畔练习伸展和手臂和腿部摆动。接下来，让游泳者进入游泳池，执行一个标准的热身方案，包括前爬行游泳800米的光强度。热身结束后，让游泳者在池畔休息10分钟。在休息时间结束时，通过 L 形扁平铁棒将称重传感器连接到启动块，用于系绳游泳测量，然后将无弹性绳索的一端连接到称重传感器。使用定制设计的皮带，该皮带将绳索连接到两个臀部，这样腿部踢不会干扰力测量，并将绳索的另一端连接到游泳者。然后确定在测量系统中以最小张力水平保持游泳者身体所需的负载。当游泳者准备好后，向游泳者发出信号，在水中开始试验，监测游泳者，并在30秒的测试中提供口头鼓励。在试验结束时，示意游泳者结束测试，然后将游泳者从没有弹性的绳索上分离。指示游泳者执行标准冷却方案，包括以光强度前爬行游泳，让游泳者在池畔休息30分钟。在休息时间结束时，将游泳者重新绑在无弹性的绳索上，并示意游泳者开始第二次全绳游泳试验。在试验结束时，信号游泳者停止测试，并执行第二个标准冷却方案，包括前爬行游泳在光强度。在冷却结束时，让游泳者离开游泳池。对于增量系绳游泳测试，在计算测试期间抵抗游泳者向前位移的负载后，验证计算机与设备软件中的自动便携式代谢单元之间的通信链路。打开装置电源，使设备预热 45 分钟，并确认电池已充满电。然后输入主题数据、环境温度和湿度。为使游泳者做好增量测试的准备，请给游泳者安装面罩和浮潜。指示游泳者在池畔休息，收集基线气体交换和通风数据。10分钟后，指示游泳者执行一个标准热身程序，包括使用浮潜系统的光强度前爬行游泳，以确保熟悉。在预热结束时，将无弹性绳索拆下到皮带上，将绳索的另一端连接到装载系统。系好游泳者腰部的腰带。指示游泳者进入游泳池，并使用池底的两个标记作为参考点，以便他们在泳池内保持相对固定的位置。当游泳者准备好时，发出信号开始测试，并在试验期间监控游泳。有研究助理经验，在监测这种类型的测试，应举行气体分析单位在池畔，而不妨碍游泳者流离失所或提升游泳者的头部。此测试需要同步工作，包括更改负载、鼓励游泳者、监测水中的位置以及正在收集的数据。在试用期间，在计时 60 秒阶段时增加负载。当游泳者不再能够保持必要的位置，尽管强烈的口头鼓励，终止测试，并记录时间限制运动容忍度。使用运动容差限制的时间计算完成的阶段，并记录每个阶段的负载和峰值负载。然后将游泳者从无弹性绳索上分离出来，并指示游泳者在离开游泳池之前执行标准冷却方案，包括以低到中等强度前爬行游泳。增量运动测试的初始负载设定为超过游泳者在开始全面游泳之前保持身体对齐所需的负载，在全游和初始全泳期间测量的平均力之间的 30% 差值。然后，每60秒阶段增加0.7公斤的负荷。游泳者运动耐受性的限制发生在576秒后的第10阶段。当在基线和渐进式运动测试的锻炼部分收集的呼吸吸氧数据平均为连续9秒的垃圾箱时，最高的三分滚动平均值为每分钟3.44升，吸氧负荷坡度为每公斤261毫升。在快速渐进式运动测试期间，动脉血二氧化碳和最终潮汐二氧化碳的减少至少两分钟或两分以上，在此期间，工作和代谢率继续上升。对于这位游泳者来说，由于厌氧通路对能量需求的贡献增加，气体交换和通气反应的这些明显变化的代谢率分别出现在吸氧峰值的75%和86%。管理测试的技术人员必须协调，因为需要精确的时间，游泳者必须具有高度的积极性和熟悉系绳游泳的新方面。由于对工作线性增加的代谢反应不是线性的，因此这种类型的测试允许识别定义运动员非线性的断点。

Research

JoVE Journal

Biochemistry

An Optimized Protocol for Electrophoretic Mobility Shift Assay Using Infrared Fluorescent Dye-labeled Oligonucleotides

一种优化的协议电泳迁移分析使用红外荧光染料标记的寡核苷酸

Electrophoretic Mobility Shift Assays (EMSA) are an instrumental tool to characterize the interactions between proteins and their target DNA sequences. ...

科研

生物化学

A Tailored HPLC Purification Protocol That Yields High-purity Amyloid Beta 42 and Amyloid Beta 40 Peptides, Capable of Oligomer Formation

一个定制HPLC纯化协议，产量高纯度淀粉样蛋白β42和贝塔淀粉样蛋白肽40，有能力寡聚物形成的

Amyloidogenic peptides such as the Alzheimer's disease-implicated Amyloid beta (A&#946;), can present a significant challenge when trying to obtain high ...

Hydrolysis of a Ni-Schiff-Base Complex Using Conditions Suitable for Retention of Acid-labile Protecting Groups

镍 - 席夫碱的水解复杂环境条件适宜酸不稳定保护基团的保留

Unnatural amino acids, amino acids containing side-chain functionalities not commonly seen in nature, are increasingly found in synthetic peptide ...

Optimized Protocol for the Extraction of Proteins from the Human Mitral Valve

从人体二尖瓣提取蛋白质的优化方案

Analysis of the cellular proteome can help to elucidate the molecular mechanisms underlying diseases due to the development of technologies that permit ...

Measuring Biomolecular DSC Profiles with Thermolabile Ligands to Rapidly Characterize Folding and Binding Interactions

用耐热配体测量生物分子 DSC 剖面快速表征折叠和结合的相互作用

Differential scanning calorimetry (DSC) is a powerful technique for quantifying thermodynamic parameters governing biomolecular folding and binding ...

Improved Protocol for Chromatin Immunoprecipitation from Mouse Skeletal Muscle

改进的小鼠骨骼肌染色质沉淀的协议

We describe an efficient and reproducible protocol for the preparation of chromatin from adult mouse skeletal muscle, a physically resistant tissue with a ...

A Western Blotting Protocol for Small Numbers of Hematopoietic Stem Cells

一种用于小数目造血干细胞的西方印迹协议

Hematopoietic stem cells (HSCs) are rare cells, with the mouse bone marrow containing only ~25,000 phenotypic long term repopulating HSCs. A Western ...

Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays

Detergent-assisted Reconstitution of Recombinant Drosophila Atlastin into Liposomes for Lipid-mixing Assays

洗涤剂辅助重组重组果蝇阿拉他素到脂质混合测定脂质体

Membrane fusion is a crucial process in the eukaryotic cell. Specialized proteins are necessary to catalyze fusion. Atlastins are endoplasmic reticulum ...

Stepwise Dosing Protocol for Increased Throughput in Label-Free Impedance-Based GPCR Assays

基于无标签阻抗的 GPCR 检测中用于提高吞吐量的逐步加量协议

Label-free impedance-based assays are increasingly used to non-invasively study ligand-induced GPCR activation in cell culture experiments. The approach ...

Assessment of Cellular Oxidation using a Subcellular Compartment-Specific Redox-Sensitive Green Fluorescent Protein

使用亚细胞室特异性红氧敏感绿色荧光蛋白评估细胞氧化

Measuring the intracellular oxidation/reduction balance provides an overview of the physiological and/or pathophysiological redox status of an organism. ...