Name: taste exam: a brief and validated test
Uploaded: 2018-08-17T12:30:00.000+00:00
Duration: 7 min 10 s
Description: Watch this Scientific Journal Video about Taste Exam: A Brief and Validated Test at JoVE.com

来自国立卫生研究院的奖项支持这项研究 (R01DC013588 到 nac、R21DC013886 和 DRR, 以及 NIDCD 行政研究补充, 以促进临床医生科学家在化学感受研究中的出现)。我们收集了部分从 OD018125 的 NIH 基金购买的设备的基因型数据。

宾夕法尼亚大学机构审查委员会批准了这项议定书。如果他们18岁以下或怀孕, 我们就排除了这些科目。
1. 口腔评价: 疾病评估和乳突鉴定
<ol><li>指示主语张开嘴。</li>
	<li>
使用光源, 如手电筒或前照灯, 照亮口腔和检查该地区的七个子网站 (舌, 口, 颊粘膜, 唇粘膜, 牙龈, 硬腭, 和 retromolar 三角)。
	<ol>
<li>可视化舌头的背表面。指导主体抬起舌头, 检查腹舌表面和口底, 确保将检查向后至磨牙。</li>
		<li>使用舌抑制剂, lateralize 的脸颊, 以可视化颊粘膜, 以及侧牙龈两侧周围的上下牙齿。</li>
		<li>通过抬起上下唇, 使检查前方, 以可视化唇粘膜和前牙龈的表面。</li>
		<li>最后, 形象化的硬腭和 retromolar 三角。</li>
		<li>注意病灶、擦伤、肿块或炎症症状。</li>
		<li>再一次, 问这个问题, 张开嘴巴, 伸出舌头。</li>
		<li>使用光源来可视化舌头的背表面。</li>
		<li>确定 FP 的存在或不存在,如平滑的舌面</li>
	</ol>
</li>
	<li>在进行味觉测试之前, 注意口腔检查的结果。如果调查人员在医学背景下进行这种味觉测试, 意想不到的发现会促使进一步的工作。</li>
</ol>2. 心理生理味觉测试
注: 下面的网页还提供了关于心理测试的资源和描述: https://osf.io/hn87s/。
<ol><li>
Tastant 准备
	<ol>
<li>按以下指示准备解决方案。使用容积瓶使每个解决方案, 以确保浓度精度为 0.0002 m. 用超纯水溶解样品。为研究目标量身定做化合物的选择。这里包含的化合物就是一个例子。<ol>
<li>地那铵苯甲酸酯 (苦味): 在1升水中溶解2.228 克苯甲酸地那铵, 以制备 4.99 x 10-3米地那铵苯甲酸酯的库存溶液。将此库存溶液的180µL 添加到500毫升容积烧瓶中。加入水, 使体积到500毫升, 产生一个溶液, 最终浓度为1.8 µM。</li>
			<li>ptc (苦涩): 在500毫升容量瓶中放置0.0135 克 ptc。加水使体积增加到500毫升。PTC 很难溶解, 所以在烧瓶中放置一个搅拌棒, 并将溶液加热到70摄氏度的热板上。使用搅拌杆混合溶液, 直到所有溶质溶解 (~ 15 分钟)。这将产生一个最终浓度为180µM 的解决方案。</li>
			<li>奎宁 (苦味): 将0.011 克奎宁盐酸盐放入500毫升容积瓶中。加入水, 使体积到500毫升, 产生一个溶液, 最终浓度为56µM。</li>
			<li>氯化钠 (咸): 在500毫升容积的烧瓶中放置7.5 克氯化钠。加入水, 使体积到500毫升, 产生一个解决方案, 最终浓度为 0.25 m。</li>
			<li>蔗糖 (甜): 在500毫升容积烧瓶中放置60克蔗糖。加入水, 使体积到500毫升, 产生一个解决方案, 最终浓度为 0.35 m。</li>
		</ol>
</li>
		<li>商店的口味解决方案在4摄氏度。一些常用的味觉化合物是光敏的, 调查人员应该用箔或其他材料包装, 以减少暴露在光线中。</li>
		<li>要确定解决方案准备中的常见错误, 请用旧的解决方案和一个新的解决方案填充一个品尝杯。品尝每个解决方案以验证它们在强度上是否相同。</li>
		<li>包括水作为控制解决方案, 提出在第一位验证对象了解测试程序。与每个 tastant 和控制 tastant 两次, 注意避免连续呈现相同的 tastant。例如, 要测试五个唯一的 tastants, 请在调查表中显示十二个样本 (见 2.2): 五 tastants 和水, 每两次提出。整除5毫升的水进入单个的玻璃闪烁瓶。标签的瓶子帽与一个深蓝色的标签轴承的数字1。</li>
		<li>对每个 tastant 重复此过程。标签瓶帽与圆形贴纸按顺序显示和一些颜色代码, 示例详细如下;将药瓶标签与味觉调查表上的标签匹配 (见 2.2)。 水 (深蓝色) 奎宁 (浅蓝色) NaCl (绿色) PTC (黄色) 蔗糖 (橙) 地那铵苯甲酸酯 (红) NaCl (深蓝色) 地那铵苯甲酸酯 (浅蓝色) 水 (绿色) 奎宁 (黄色) 蔗糖 (橙) PTC (红色)</li>
		<li>将样品包装成两个盒子, 分别在标签为 "方框 A" 和 "方框 B" 的方框中取样1–6和 7–12 (图 1);其他的包装策略是可能的。</li>
	</ol>
</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56705/56705fig1v2.jpg"> 图 1: 味觉套件.受试者使用该套件来对各种颜色编码的 tastants 的口感强度和质量进行率。方框 A 包含示例 1–6, 方框 B 包含7–12。<a href="/files/ftp_upload/56705/56705fig1v2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="2"><li>
品味问卷
	<ol>
<li>使用类别刻度来准备味觉调查表, 以评定味觉强度和确定每个 tastant 的口感质量的强制选择 (图 2)。</li>
		<li>在味觉调查表中, 在适当的样品旁边放置适当颜色和编号的圆形标签 (见 2.1.6)。</li>
	</ol>
</li>
</ol><img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56705/56705fig2.jpg"> 图 2: 味觉调查表条目, 包括强度分级的类别刻度和 tastant 质量的强制选择响应.味觉调查表将包括每个测试的彩色编码 tastants 的一个条目。<a href="/files/ftp_upload/56705/56705fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="3"><li>
味觉测试管理
	<ol>
<li>为主题提供方框 A、方框 B、一瓶水、空杯子、钢笔和笔纸味觉问卷, 其中包含12个样本的条目。在任何特定的研究期间, 使用相同牌子的瓶装水。作为一份纸质问卷的替代品, 将材料放入在线调查软件中, 并通过平板电脑或台式机/笔记本电脑进行管理。示例模板可在 https://osf.io/hn87s/中使用。</li>
		<li>指导主题, 他们将被要求对每 tastant 的强度和质量 (如咸, 酸, 苦, 甜, 或没有味道) 进行率。此外, 告知主题, 他们可能不会经历所有的品质。</li>
		<li>解释测试程序, 如下所示: 用清水冲洗两次嘴, 并在所提供的杯子里吐出来。把所有样品1倒入你的嘴里, 把它放在那里5秒钟, 然后把溶液吐到杯子里。不要漱口或吞下溶液。圆形的13条垂直线与样品的强度相对应, 在0到12的尺度上, 从 "无强度" 到 "极强", 选择单一品质来描述味道。然后, 用清水冲洗你的嘴两次, 然后再继续下一个样品。</li>
		<li>观察主题品尝和评分样本 1 (水)。如果评级偏离 "根本没有强度" 和 "无风味", 则在允许测试继续之前, 重复调查表的说明。</li>
		<li>检查完成的调查表是否完整。</li>
		<li>评分强度评分的比例为0到12从垂直线, 主题盘旋。平均两个 tastant 的强度等级;此值将用于分析。</li>
	</ol>
</li>
</ol>3. 基因型
<ol><li>使用唾液 DNA 收集工具收集每个主题的唾液。</li>
	<li>按照制造商的指示, 从样品中纯化基因组 DNA。</li>
	<li>用 SNP 基因分型测定 (rs713598,rs1726866, rs10246939) 测定 TAS2R38基因。</li>
	<li>用 SNP 基因分型测定 (rs10772420)36测定TAS2R19基因。</li>
</ol>4. 基因型-表位验证
<ol><li>在下面的网页中, 查看来自800多个主题 genotyped 的可用池控制数据 (rs713598、rs1726866、rs10246939) 和TAS2R19 (rs10772420): https://carayata.shinyapps.io/TasteBoxplots/。</li>
	<li>基于一个主题的TAS2R38基因型, 比较他或她的心理味觉反应的苦复方 PTC 与规范的个体的同一基因型。反应应匹配;然而, 在罕见的情况下, TAS2R38基因型并不完全预测 PTC 感官结果35。</li>
	<li>如果反应显示出显著的差异, 比较的主题的味觉反应的奎宁与规范的个人相同的TAS2R19基因型。对奎宁强度和基因型的反应应匹配36。如果所有的味觉结果都不能与基因型相对应, 那么这个主题 (a) 就可能不理解任务 (b) 是否提供了虚假评级或伪装, 或者 (c) 调查人员的数据收集错误。</li>
	<li>识别数据点的异常值, 并可能将它们排除在分析中 (图 3)。感官结果与客观基因分型的对应性验证了心理测试程序的可靠性。</li>
</ol>

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A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Putative+mammalian+taste+receptors:+a+class+of+taste-specific+GPCRs+with+distinct+topographic+selectivity.">Putative mammalian taste receptors: a class of taste-specific GPCRs with distinct topographic selectivity.</a> Cell. 96 (4), 541-551 (1999).</li><li>Laffitte, A., Neiers, F., Briand, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+roles+of+the+sweet+taste+receptor+in+oral+and+extraoral+tissues.">Functional roles of the sweet taste receptor in oral and extraoral tissues.</a> Current Opinion in Clinical Nutrition and Metabolic. 17 (4), 379-385 (2014).</li><li>An, S. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tas2r+activation+promotes+airway+smooth+muscle+relaxation+despite+beta2-adrenergic+receptor+tachyphylaxis.">Tas2r activation promotes airway smooth muscle relaxation despite beta2-adrenergic receptor tachyphylaxis.</a> American Journal of Physiology-Lung Cellular and Molecular. , (2012).</li><li>Fox, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+chemical+composition+and+taste.">The relationship between chemical composition and taste.</a> Science. 74, 607 (1931).</li><li>Fox, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+relationship+between+chemical+constitution+and+taste.">The relationship between chemical constitution and taste.</a> Proceedings of the National Academy of Sciences USA. 18, 115-120 (1932).</li><li>Bufe, B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+molecular+basis+of+individual+differences+in+phenylthiocarbamide+and+propylthiouracil+bitterness+perception.">The molecular basis of individual differences in phenylthiocarbamide and propylthiouracil bitterness perception.</a> Current Biol.ogy. 15 (4), 322-327 (2005).</li><li>Reed, D. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+perception+of+quinine+taste+intensity+is+associated+with+common+genetic+variants+in+a+bitter+receptor+cluster+on+chromosome+12.">The perception of quinine taste intensity is associated with common genetic variants in a bitter receptor cluster on chromosome 12.</a> Human Molecular Genetics. 19 (21), 4278-4285 (2010).</li><li>Bobowski, N., Reed, D. R., Mennella, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Variation+in+the+TAS2R31+bitter+taste+receptor+gene+relates+to+liking+for+the+nonnutritive+sweetener+Acesulfame-K+among+children+and+adults.">Variation in the TAS2R31 bitter taste receptor gene relates to liking for the nonnutritive sweetener Acesulfame-K among children and adults.</a> Science Reports. 6, 39135 (2016).</li><li>Allen, A. L., McGeary, J. E., Knopik, V. S., Hayes, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bitterness+of+the+non-nutritive+sweetener+acesulfame+potassium+varies+with+polymorphisms+in+TAS2R9+and+TAS2R31.">Bitterness of the non-nutritive sweetener acesulfame potassium varies with polymorphisms in TAS2R9 and TAS2R31.</a> Chemical Senses. 38 (5), 379-389 (2013).</li><li>Roudnitzky, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genomic,+genetic,+and+functional+dissection+of+bitter+taste+responses+to+artificial+sweeteners.">Genomic, genetic, and functional dissection of bitter taste responses to artificial sweeteners.</a> Human Molecular Genetics. 20 (17), 3437-3449 (2011).</li><li>Guo, S. W., Reed, D. 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NAC 和 DRR 是共同发明者在被审查的专利 (治疗和诊断为呼吸道传染 61/697652, WO2013112865)。

这种方法的意义在于它采用了多学科的方法, 具有医学 (口试)、实验心理学 (味觉测试) 和遗传学 (验证步骤) 的特点。味觉信息可能会发展成为一种诊断和预后的工具, 因为味觉提供了一个窗口, 在身体其他地方的蛋白质功能。从实验心理学的角度看, 增加一个简单的考试可以识别不适合研究规范味觉功能的科目。从遗传学的角度来看, 这些程序提供了一种简单的方法来研究容易重现的基因型-表观关系。
测量人的味觉有几个关键的特征是无形的, 但重要的, 包括帮助主体感到轻松和面向的任务, 特别是在医疗设置, 保持程序短, 所以关注的主题不动摇。同样重要的是, 如果受试者看起来不舒服, 并且麻烦地拍摄他们的担忧, 比如关于测试刺激的性质的话。受试者通常会确信, 大多数的测试刺激都在食物中,例如盐和糖。这个过程虽然简单, 但有很大的局限性。虽然口试是调查人员接受医学训练的例行程序, 但那些有实验心理学或遗传学训练的人在识别口腔疾病方面可能不那么容易。另一个限制是评分表, 虽然没有事先培训的受试者容易理解, 但可能会模糊个人或个人群体之间的差异41。最后, 在数据统计处理中, 如何处理与基因型和表型不匹配的主题的指导还未编入简单的规则,例如, 删除那些未能达到一定标准的科目。
展望未来的应用, 味觉考试可能成为医学的常规部分, 如视力和听力测试, 这将增加我们对味觉与人类疾病和福祉的关系的理解, 并将使我们能够改进这个简单的测试。

人类味觉知觉的衡量方法既可以是医疗保健的一部分, 也是生物医学研究的目标, 但与听觉和视力相比, 味觉却很少受到重视 (表 1)。从医学的角度来看, 当临床医生评估病人抱怨的味觉损失, 在大多数情况下, 实际损失的气味1, 这导致解雇的味觉损失作为一个罕见的, 往往是无效的提出投诉。味觉畸变 (dysgeusia) 是更常见的, 经常出现的副作用的药物或周围神经损伤2,3, 但两种形式都没有有效的治疗 (除了停止用药)。临床医生也忽略了味觉的丧失, 因为它迄今对自身的诊断和预后价值甚少。然而, 虽然味觉的测量是一个回水, 它现在可能进入主流医学与复兴的历史欣赏, 味觉可能是诊断或预后工具4,5。例如, 苦味知觉可以预测免疫功能6或患者愿意服用药物7。然而, 生物医学研究人员基本上忽略了味觉。这种疏忽可能在一定程度上反映了这样一个事实: 在理解这个感官系统的早期进展有它的根源在实验心理学8, 一个领域与那些在医学可能是相对地不熟悉的。此外, 对味觉的重新兴趣带来了标准化的味觉方法9 , 建立在早期的方法10, 而全面是漫长的, 不适合临床设置。最后, 对味觉措施的信心可能是微弱的, 因为受试者报告他们自己的经验和证实他们的观察一直是缺乏的。我们的希望是, 一个简单的措施, 调查人员或临床医生可以很容易地管理将获得在流行与两个成分。这里我们描述了一个简单的味觉检查协议, 它有三部分: 对口腔的评估, 味觉测试, 以及使用先天基因型的验证步骤。首先, 我们为这些程序提供了生物学背景, 将医学中的简单实践、实验心理学的感官测量和基因型和遗传学的反应验证结合起来。
味觉知觉开始在嘴里, 因此有效的味觉检查需要包括简要的临床评估为明显的口腔疾病, 红肿, 肿胀和其他变色。口腔含七个子体: 舌、牙龈、口底、颊粘膜、唇粘膜、硬腭、retromolar 三角。以往对人体味觉的研究集中在健康参与者或有明确疾病的人身上, 但随着味觉测试成为医学考试的常规, 重要的是记录口腔的状况作为手术的一部分。
舌头本身是一种肌肉结构, 包裹在粘膜中;点缀其背表面是乳突, 小凸起的结构, 使舌头其独特的纹理和含有味觉受体细胞。我们根据它们的形状对乳突进行分类: 蕈、丝状、叶酸和 circumvallate。蕈乳突 (FP) 位于 anterolaterally 舌上, 呈圆形, 有蘑菇状11。调查人员发表了几种有用的方法来量化 FP, 我们直接向读者提供了测量协议12、13、14、15、16的信息。叶酸的乳突, 形状像一本书的书页 (folia), 完全位于侧后舌表面11。Circumvallate 的乳突, 发现在舌底的沟终, 是大圆顶状结构包围的粘膜壁 (拉丁环, "环绕," +壁垒, "墙")11。最大量的乳突, 丝状, 长而薄, 不含味觉受体。
人的舌头解剖不同。虽然这种解剖变异的来源是未知的, 它部分地由先天遗传变异决定, 调查人员报告31% 卵双胞胎舌解剖学的一致性和60% 单卵双生子双胞胎17之间的一致性。在人群中, 乳突密度也不同, 虽然罕见, 至少有一个遗传性疾病 (家族性 dysautonomia) 导致先天性缺乏味觉乳癌18,19,20。因此, 在进行心理测试之前, 确认 FP 的存在是简要评估的一部分, 并注意到口腔疾病的舌头和证据的相对大小和颜色。
口感乳突含有刺激引发味觉感觉的感官细胞。人类能够感知至少五类的味道: 咸, 酸, 苦, 甜, 和鲜味。虽然咸, 甜, 和鲜味的味道信号的存在有价值的食物来源含有氯化钠, 葡萄糖和氨基酸, 分别, 苦味和酸味信号的存在潜在的毒素和酸的细菌分解食物, 分别和诱发厌恶行为21。咸和酸的味道是转基因通过活化离子通道发现在某些类型的味觉细胞, 虽然对盐转导的理解正在演变, 它可能需要 I 型细胞, 以及22,23。苦、甜、鲜味是由 G 蛋白偶联受体在 II. 型味觉细胞上活化而成, 每一种都能适应特定的口味。三种特殊受体亚基的 Heterodimers 传感器甜和鲜味, 而苦味化合物激活25种不同苦味受体24的组。这些苦味的受体可以反应多种苦味化合物, 一个单一的苦味化合物往往刺激超过一个受体25。尽管最近关于味觉的分子基础知识的扩展, 新的路径26和超越传统的五种口味品质 (例如, 钙27或脂肪酸28知觉) 的最新发现可能会在前面。
有至少两个令人惊讶的方面的味觉家庭的受体: 基因, 这些受体编码可以明显不同的 DNA 序列, 从而在人之间的功能, 和身体的许多组织表达这些基因21,29,30,31. 这些口外部位包括脑、甲状腺、上、下呼吸道和胃肠道, 其中许多人21、29、30、31。当口味感受器官在这些地点不参与口味知觉在传统感觉, 他们可能感觉地方化学环境29,32。例如, 上呼吸道纤毛上皮表达苦味受体 T2R38 (苦味受体 38), 它响应细菌产生的化合物, 并影响先天免疫反应32, 如增加黏膜纤毛间隙和抗微生物肽和一氧化氮的水平。这一发现对慢性鼻窦炎、慢性细菌感染疾病和上呼吸道和鼻窦炎症有医学意义。
与我们这里描述的味觉检查特别相关的是, T2R38 苦味受体, 编码的TAS2R38基因, 表现出遗传变异, 因此变化的味觉敏感性。对苦味化合物 phenylthiocarbamide (PTC) 的知觉差异首先由化学家亚瑟狐狸33描述;后来这个化合物被确定为 T2R38 受体34的激动剂。个体差异产生于 DNA 序列的TAS2R38基因, 其中有三单核苷酸多态性, 每一个产生氨基酸替代品 (A49P, A262V 和 I296V;A: 丙氨酸, P: 脯氨酸, V: 缬氨酸, I。两个常见的单体结果, PAV 和 AVI, 与 PAV/PAV 个人高度敏感的 PTC ("品酒师"), avi/avi 个人是相对不敏感 ("非品酒师"), 和异型的 AVI/PAV 个人在他们的灵敏度更可变35. 有更多的基因变异的例子影响苦味,例如, TAS2R19基因编码的味觉受体 T2R19, 同样表现出遗传变异和对苦味化合物的不同口味敏感性。奎宁36同样, TAS2R31的变异也会影响到高效甜味剂37、38、39的感知苦味。
在这里, 我们描述了一个快速的方法来表征病人的味觉, 利用高产量的协议, 在临床医学, 实验心理学和遗传学。

<table><tbody><tr><td>Disposable diagnostic penlight</td><td>Primacare</td><td>DL-9223</td><td></td></tr><tr><td>UltraLite Pro headlight</td><td>Integra LifeSciences</td><td>AX2100BIF</td><td></td></tr><tr><td>Millipore Q-Gard 2 water purification system</td><td>EMB Millipore</td><td>QGARD00D2</td><td></td></tr><tr><td>Denatonium benzoate&amp;nbsp;</td><td>Sigma Aldrich</td><td>D5765</td><td></td></tr><tr><td>Phenylthiocarbamide</td><td>Sigma Aldrich</td><td>P7629</td><td></td></tr><tr><td>Quinine hydrochloride dihydrate</td><td>Sigma Aldrich</td><td>Q1125</td><td></td></tr><tr><td>Sodium Chloride</td><td>Sigma Aldrich</td><td>S1679</td><td></td></tr><tr><td>Sucrose</td><td>Sigma Aldrich</td><td>S0389</td><td></td></tr><tr><td>Glass scintillation vials</td><td>Thomas Scientific</td><td>1230L59</td><td>Same as Wheaton catalog no. 986580</td></tr><tr><td>Oragene Discover OGR-500 DNA collection kit</td><td>DNA Genotek</td><td>OGR-500</td><td></td></tr><tr><td>prepIT L2P Protocol reagents</td><td>DNA Genotek</td><td>PT-L2P-5</td><td></td></tr><tr><td>&lt;em&gt;rs713598 &lt;/em&gt;TaqMan SNP genotyping assay</td><td>ThermoFisher Scientific</td><td>C___8876467_10</td><td></td></tr><tr><td>&lt;em&gt;rs1726866 &lt;/em&gt;TaqMan SNP genotyping assay</td><td>ThermoFisher Scientific</td><td>C___9506827_10</td><td></td></tr><tr><td>&lt;em&gt;rs10246939 &lt;/em&gt;TaqMan SNP genotyping assay</td><td>ThermoFisher Scientific</td><td>C___9506826_10</td><td></td></tr><tr><td>&lt;em&gt;rs10772420 &lt;/em&gt;TaqMan SNP genotyping assay</td><td>ThermoFisher Scientific</td><td>C___1317426_10</td><td></td></tr></tbody></table>

taste exam: a brief and validated test

味觉在医学和生物医学研究中的日益重要, 以及对其遗传基础的新认识, 促使我们在两方面补充了经典的味觉测试方法。首先, 我们解释如何做一个简短的评价口腔, 包括舌头, 以确保味觉乳突存在, 并注意到有关疾病的证据。其次, 通过比较感知苦味强度和先天受体基因型的报告, 利用遗传学来验证味觉测试数据。基因型客观测度与味觉体验主观报告之间的不一致可以识别数据收集错误、分心的主题或那些不理解或遵循指令的人。我们的期望是快速有效的味觉测试可以说服研究人员和临床医生定期评估味觉, 使味觉测试与听力和视力测试一样普遍。最后, 由于许多组织的身体表达味觉受体, 味觉反应可能提供一个代理的组织敏感性在其他地方的身体, 从而, 作为一个快速的, 护理点测试指导诊断和研究工具, 评估味觉受体蛋白质功能。

从味觉测试的结果已汇集为所有被评估的科目 (n = 840), 并提出后, 分离的基因型。完整的数据集可在 https://carayata.shinyapps.io/TasteBoxplots/中访问, 可以对评估的每个 tastant 和TAS2R38和TAS2R19基因型进行审查。结果证实了TAS2R38受体基因型 (图 3) 中 PTC 的知觉味觉差异的存在。PTC 强度的评分在TAS2R38基因型 (avi/avi, 0.86) 有显著差异;AVI/PAV, 6.95;PAV/PAV, 8.18;单向方差分析, p < 0.0001)。在TAS2R19基因型 (a: 3.77, 奎宁强度的结果也有显著差异;A: G, 3.08;g: g, 2.26;单向方差分析, p < 0.0001)。
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56705/56705fig3.jpg"> 图 3: PTC 味觉问卷结果TAS2R38基因型。味觉问卷可以用来隔离个体的TAS2R38基因型基于 PTC 苦味强度评分的类别规模 (* p < 0.0001)。在这里, 我们看到了一些异常点, 位于这个箱线图的栅栏 ("胡须") 外的数据点数 (例如, 在上面四分位以上的四分位范围的1.5 倍以上和低四分位的波纹管)<a href="/files/ftp_upload/56705/56705fig3large.jpg" target="_blank">请点击这里查看这个更大的版本图.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>度量</td>
			<td>味道</td>
			<td>听到</td>
			<td>视觉</td>
			<td>听觉/味觉</td>
			<td>视觉/品味</td>
		</tr>
<tr>
<td>Pubmed</td>
			<td>36302</td>
			<td>123101</td>
			<td>171522</td>
			<td>3.39</td>
			<td>4.72</td>
		</tr>
<tr>
<td>NIH 记者</td>
			<td>6144</td>
			<td>23873</td>
			<td>54858</td>
			<td>3.89</td>
			<td>8.93</td>
		</tr>
<tr>
<td colspan="6">我们进行了PubMed和NIH 记者的文本搜索, 以某种意义上的关键字。PubMed是一个发布实验结果的在线数据库, NIH 的记者是一个在线数据库, 列出了由国立卫生研究院资助的研究项目。"听觉/味觉" 栏目中的价值观显示出一个比率,也就是说, 与味觉相比, 引用听力的出版物有三倍多。我们访问了下面的网址在2018年1月31日在上午10点 EST。</td>
		</tr>
<tr>
<td colspan="5">https://www.ncbi.nlm.nih.gov/pubmed/</td>
			<td></td>
		</tr>
<tr>
<td colspan="5">https://projectreporter.nih.gov/reporter.cfm</td>
			<td></td>
		</tr>
</tbody></table>表 1: 使用 "味觉" 与 "听觉" 和 "视觉" 作为关键词识别的记录。

Watch this Scientific Journal Video about Taste Exam: A Brief and Validated Test at JoVE.com

Taste Exam: A Brief and Validated Test

味觉考试: 一个简短的验证测试

该协议测量人的味觉反应, 包括一个简短的解剖评估, 短的味觉测试, 和一个验证方法使用的主题的感觉和味觉受体基因型。

The emerging importance of taste in medicine and biomedical research, and new knowledge about its genetic underpinnings, has motivated us to supplement ...

Universidad San Sebastian

Research

JoVE Journal

Medicine

16.2K 次观看.  Perelman School of Medicine at the University of Pennsylvania. 该协议测量人的味觉反应, 包括一个简短的解剖评估, 短的味觉测试, 和一个验证方法使用的主题的感觉和味觉受体基因型。

视频: Taste Exam: A Brief and Validated Test

Examination of Rapid Dopamine Dynamics with Fast Scan Cyclic Voltammetry During Intra-oral Tastant Administration in Awake Rats

Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. Fast scan cyclic voltammetry (FSCV) is an electrochemical technique with high spatial and temporal (sub-second) resolution that has been utilized to examine phasic DA release in several types of preparations. In vitro experiments in single-cells and brain slices and in vivo experiments in anesthetized rodents have been used to identify mechanisms that mediate dopamine release and uptake under normal conditions and in disease models. Over the last 20 years, in vivo FSCV experiments in awake, freely moving rodents have also provided insight of dopaminergic mechanisms in reward processing and reward learning. One major advantage of the awake, freely moving preparation is the ability to examine rapid DA fluctuations that are time-locked to specific behavioral events or to reward or cue presentation. However, one limitation of combined behavior and voltammetry experiments is the difficulty of dissociating DA effects that are specific to primary rewarding or aversive stimuli from co-occurring DA fluctuations that mediate reward-directed or other motor behaviors. Here, we describe a combined method using in vivo FSCV and intra-oral infusion in an awake rat to directly investigate DA responses to oral tastants. In these experiments, oral tastants are infused directly to the palate of the rat &#8211; bypassing reward-directed behavior and voluntary drinking behavior &#8211; allowing for direct examination of DA responses to tastant stimuli.

Rapid fluctuations in extracellular dopamine (DA) mediate both reward processing and motivated behavior in mammals. This manuscript describes the combined use of fast scan cyclic voltammetry (FSCV) and intra-oral tastant administration to determine how tastants alter rapid dopamine release in awake, freely moving rats.

考试多巴胺快速与动态快速扫描循环伏安法在口内呈味物质管理清醒大鼠

Rapid, phasic dopamine (DA) release in the mammalian brain plays a critical role in reward processing, reinforcement learning, and motivational control. ...

科研

行为学

Protocol for Data Collection and Analysis Applied to Automated Facial Expression Analysis Technology and Temporal Analysis for Sensory Evaluation

We demonstrate a method for capturing emotional response to beverages and liquefied foods in a sensory evaluation laboratory using automated facial expression analysis (AFEA) software. Additionally, we demonstrate a method for extracting relevant emotional data output and plotting the emotional response of a population over a specified time frame. By time pairing each participant's treatment response to a control stimulus (baseline), the overall emotional response over time and across multiple participants can be quantified. AFEA is a prospective analytical tool for assessing unbiased response to food and beverages. At present, most research has mainly focused on beverages. Methodologies and analyses have not yet been standardized for the application of AFEA to beverages and foods; however, a consistent standard methodology is needed. Optimizing video capture procedures and resulting video quality aids in a successful collection of emotional response to foods. Furthermore, the methodology of data analysis is novel for extracting the pertinent data relevant to the emotional response. The combinations of video capture optimization and data analysis will aid in standardizing the protocol for automated facial expression analysis and interpretation of emotional response data.

A protocol for capturing and statistically analyzing emotional response of a population to beverages and liquefied foods in a sensory evaluation laboratory using automated facial expression analysis software is described.

协议进行数据采集和分析应用到自动人脸表情分析技术和时序分析感官评价

We demonstrate a method for capturing emotional response to beverages and liquefied foods in a sensory evaluation laboratory using automated facial ...

Psychophysical Tracking Method to Measure Taste Preferences in Children and Adults

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to adulthood. The method, which is identical for children, adolescents, and adults, is of short duration (&#60; 15 min), does not rely on sustained attention or place demands on memory (which would yield spurious age differences), and minimizes the impact of language development, making this method amenable to the cognitive limitations of pediatric populations. In this whole-mouth tasting method, subjects are asked to taste (without swallowing) pairs of solutions of different sucrose concentrations and to point to the solution they prefer. Each subsequent pair contains the participant&#39;s preceding preferred concentration and an adjacent stimulus concentration. The procedure continues until the subject chooses either a given concentration of sucrose when paired with both a higher and a lower concentration, or the highest or lowest concentration two consecutive times. Subjects are prevented from reaching response criteria on the basis of first or second position bias by the two-series design of the method, which counterbalances the order of solution presentation within each pair between the series (the weaker concentration is presented first in Series 1, second in Series 2). The geometric mean of the two sucrose concentrations chosen in Series 1 and 2 is an estimate of the participant&#39;s most preferred level of sucrose. Sucrose preference as determined with this laboratory-based measure has been shown to be associated with preference for sugars in foods and beverages and with taste receptor genotype, family history of alcoholism, and race/ethnicity, as well as depressive symptomatology among pediatric populations. The method has real-world relevance and has been applied to determine most preferred level of other tastes (e.g., salt), making it a valuable psychophysical tool.

Sweet taste has powerful hedonic appeal among people of all ages, particularly children. Described herein is a reliable and valid method that can be used to determine the level of sweetness most preferred, making it a valuable psychophysical tool for scientists.

心理跟踪方法来衡量儿童和成人的口味偏好

The Monell two-series, forced-choice, paired-comparison tracking method provides a reliable measure of sweet taste preferences from childhood to ...

Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty acid receptors housed on taste cells. Previous research has shown that an individual&#39;s oral sensitivity to fatty acid, specifically oleic acid (C18:1) is associated with body mass index (BMI), dietary fat consumption, and the ability to identify fat in foods. We have developed a reliable and reproducible method to assess oral chemoreception of fatty acids, using a milk and C18:1 emulsion, together with an ascending forced choice triangle procedure. In parallel, a food matrix has been developed to assess an individual&#39;s ability to perceive fat, in addition to a simple method to assess fatty food liking. As an added measure tongue photography is used to assess papillae density, with higher density often being associated with increased taste sensitivity.

Oral chemoreception of fatty acids and the association with diet and fatty food preferences may enable the identification of mechanisms involved with the development of obesity and why dietary changes may be difficult for many individuals.

测量口腔脂肪酸阈值，感知脂肪，脂肪食物喜欢的，和乳头密度的人类

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty ...

神经科学

Taste Preference Assay for Adult Drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

口味偏好测定成人<em&gt;果蝇</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

New Methods to Study Gustatory Coding

The sense of taste allows animals to detect chemicals in the environment, giving rise to behaviors critical for survival. When Gustatory Receptor Neurons (GRNs) detect tastant molecules, they encode information about the identity and concentration of the tastant as patterns of electrical activity that then propagate to follower neurons in the brain. These patterns constitute internal representations of the tastant, which then allow the animal to select actions and form memories. The use of relatively simple animal models has been a powerful tool to study basic principles in sensory coding. Here, we propose three new methods to study gustatory coding using the moth Manduca sexta. First, we present a dissection procedure for exposing the maxillary nerves and the subesophageal zone (SEZ), allowing recording of the activity of GRNs from their axons. Second, we describe the use of extracellular electrodes to record the activity of multiple GRNs by placing tetrode wires directly into the maxillary nerve. Third, we present a new system for delivering and monitoring, with high temporal precision, pulses of different tastants. These methods allow the characterization of neuronal responses in vivo directly from GRNs before, during and after tastants are delivered. We provide examples of voltage traces recorded from multiple GRNs, and present an example of how a spike sorting technique can be applied to the data to identify the responses of individual neurons. Finally, to validate our recording approach, we compare extracellular recordings obtained from GRNs with tetrodes to intracellular recordings obtained with sharp glass electrodes.

We present three new methods to study gustatory coding. Using a simple animal, the moth Manduca sexta (Manduca), we describe a dissection protocol, the use of extracellular tetrodes to record the activity of multiple gustatory receptor neurons, and a system for delivering and monitoring precisely timed pulses of tastants.

研究风味编码的新方法

The sense of taste allows animals to detect chemicals in the environment, giving rise to behaviors critical for survival. When Gustatory Receptor Neurons ...

A Rapid Food-Preference Assay in Drosophila

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and&#160;allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.

A Rapid Food-Preference Assay in Drosophila

We present a protocol for a two-choice feeding assay for flies. This feeding assay is fast and easy to run and is suitable not only for small-scale laboratory research, but also for high-throughput behavioral screens in flies.

果蝇的快速食物偏好分析

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate ...

Whole-Mount Staining, Visualization, and Analysis of Fungiform, Circumvallate, and Palate Taste Buds

Taste buds are collections of taste-transducing cells specialized to detect subsets of chemical stimuli in the oral cavity. These transducing cells communicate with nerve fibers that carry this information to the brain. Because taste-transducing cells continuously die and are replaced throughout adulthood, the taste-bud environment is both complex and dynamic, requiring detailed analyses of its cell types, their locations, and any physical relationships between them. Detailed analyses have been limited by tongue-tissue heterogeneity and density that have significantly reduced antibody permeability. These obstacles require sectioning protocols that result in splitting taste buds across sections so that measurements are only approximated, and cell relationships are lost. To overcome these challenges, the methods described herein involve collecting, imaging, and analyzing whole taste buds and individual terminal arbors from three taste regions: fungiform papillae, circumvallate papillae, and the palate. Collecting whole taste buds reduces bias and technical variability and can be used to report absolute numbers for features including taste-bud volume, total taste-bud innervation, transducing-cell counts, and the morphology of individual terminal arbors. To demonstrate the advantages of this method, this paper provides comparisons of taste bud and innervation volumes between fungiform and circumvallate taste buds using a general taste-bud marker and a label for all taste fibers. A workflow for the use of sparse-cell genetic labeling of taste neurons (with labeled subsets of taste-transducing cells) is also provided. This workflow analyzes the structures of individual taste-nerve arbors, cell type numbers, and the physical relationships between cells using image analysis software. Together, these workflows provide a novel approach for tissue preparation and analysis of both whole taste buds and the complete morphology of their innervating arbors.

This paper describes methods for tissue preparation, staining, and analysis of whole fungiform, circumvallate, and palate taste buds that consistently yield whole and intact taste buds (including the nerve fibers that innervate them) and maintain the relationships between structures within taste buds and the surrounding papilla.

真菌形态、环形和味觉味蕾的全镶嵌染色、可视化和分析

Taste buds are collections of taste-transducing cells specialized to detect subsets of chemical stimuli in the oral cavity. These transducing cells ...

&#181;Tongue: A Microfluidics-Based Functional Imaging Platform for the Tongue In Vivo

Intravital fluorescence microscopy is a tool used widely to study multicellular dynamics in a live animal. However, it has not been successfully used in the taste sensory organ. By integrating microfluidics into the intravital tongue imaging window, the &#181;Tongue provides reliable functional images of taste cells in vivo under controlled exposure to multiple tastants. In this paper, a detailed step-by-step procedure to utilize the &#181;Tongue system is presented. There are five subsections: preparing of tastant solutions, setting up of a microfluidic module, sample mounting, acquiring functional image data, and data analysis. Some tips and techniques to solve the practical issues that may arise when using the &#181;Tongue are also presented.

 &#181;Tongue: A Microfluidics-Based Functional Imaging Platform for the Tongue In Vivo

The article introduces the &#181;Tongue (microfluidics-on-a-tongue) device for functional taste cell imaging in vivo by integrating microfluidics into an intravital imaging window on the tongue.

μTongue:一个基于微流体的功能成像平台，用于在Viva的舌头

Intravital fluorescence microscopy is a tool used widely to study multicellular dynamics in a live animal. However, it has not been successfully used in ...

Psychophysical Tracking Method to Assess Taste Detection Thresholds in Children, Adolescents, and Adults: The Taste Detection Threshold (TDT) Test

This paper describes a two-alternative, forced-choice, staircase, tracking procedure, called the Taste Detection Threshold (TDT) test, that provides a reliable measure of sweet, salty, and umami taste detection thresholds from childhood to adulthood. Advantages of the method include procedures that are identical for children and adults, thus allowing the determination of age-related and individual differences in taste perception, if any, and tasks that can be completed in a relatively short time frame, do not rely on continuous attention or require memorization, control for subjective response biases, and minimize the impact of language development. After a 1 hour fast, participants are presented with pairs of solutions; in each pair, one solution is water, and the other solution contains varying concentrations of the tastant.
Using a whole-mouth tasting method, participants taste each solution (without swallowing and with rinsing between tastings) and then point to the solution with a taste or that tastes different from water. The concentration of the stimulus in the subsequent pair increases after a single incorrect response and decreases after two consecutive correct responses. A reversal occurs when the concentration sequence changes direction. The task is deemed completed after the occurrence of four reversals, provided there are a maximum of two dilution steps between two successive reversals, and the series of reversals do not form an ascending pattern. These additional criteria ensure greater reliability in outcomes. The TDT is then calculated as the geometric mean of the concentrations of the four reversals. This method has real-world relevance as it provides information on a dimension of taste perception that is independent of hedonics, and that can change with aging and certain disease states, making it a valuable psychophysical test.

Psychophysical Tracking Method to Assess Taste Detection Thresholds in Children, Adolescents, and Adults: The Taste Detection Threshold TDT Test

Psychophysical tools measure the functionality of the taste system for both research and health assessment purposes. This paper describes a method to measure taste detection thresholds that can determine the lowest concentration of sucrose, sodium chloride, or monosodium glutamate that can be tasted by individuals as young as 6 years.

评估儿童、青少年和成人味觉检测阈值的心理物理跟踪方法:味觉检测阈值（TDT）测试

This paper describes a two-alternative, forced-choice, staircase, tracking procedure, called the Taste Detection Threshold (TDT) test, that provides a ...