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

국립 보건원에서 선수들이이 연구 지원 (NAC, NAC를 DRR, R21DC013886 및 NIDCD 관리 연구 보충을 촉진 출현의 임상-과학자 제 드 Chemosensory 연구에에 R01DC013588). 우리는 OD018125에서 NIH 자금 부분에서 구입한 장비에서 유전자 데이터를 수집.

펜실베니아 기관 검토 위원회의 대학이이 프로토콜을 승인 했다. 우리는 그들이 18 세 미만 또는 임신 경우 과목을 제외.
1. 구강 평가: 질병 평가 및 시 식별
<ol><li>입 열을 지시 합니다.</li>
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Penlight 또는 헤드 램프와 같은 광원을 사용 하 여, 구강을 조명 하 고 지역 (혀, 입, 볼 점 막, 입술 점 막, gingiva, 하드 미각, 그리고 retromolar trigone의 바닥)의 7 개의 하위 검사.</str...

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이 방법의 중요성은 그것 기능 의학 (구두 시험), 실험 심리학 (맛 테스트), 유전학 (유효성 검사 단계)에서 다양 한 접근 방법을 사용. 맛 정보 맛 본문에 다른 단백질의 기능으로 창을 제공 하기 때문에 진단 및 예 후 도구로 개발 가능성이 높습니다. 실험 심리학 관점에서 간단한 시험의 추가 규범적인 맛 기능 연구에 적합 하지 않은 주제를 식별할 수 있습니다. 유전학 관점에서 이러한 절차는 쉽?...

의료의 일부와 생물 의학 연구의 대상이 인간의 맛 지 각의 측정 될 수 있습니다 아직 맛 청각과 비전 (표 1)에 비해 부족 한 관심을 받고 있다. 의료 관점에서 임상 환자를 평가 하는 경우 맛 감소, 대부분의 경우에서의 불평 실제 손실이입니다 냄새1, 흔히 고 종종 잘못 제시 불만으로 맛 손실의 해 고를 주도하 고 있다. 맛 왜곡 (dysgeusia)는 더 일반적이 고 자주 약물 또는 주변 신경 상해2,3, 보조 효과에서 발생 하지만 어느 형태 (이외에 약물을 멈추기) 효과적인 치료. 그것은 지금까지 자체에 작은 진단 또는 전조 값을 했다 때문에 임상 맛 손실을 무시 됩니다 또한 있다. 그러나, 비록 맛의 측정 한 침체, 그것 수 있습니다 이제 수 입력 역사적인 감사 맛 진단 또는 전조 도구4,5있을 수 있습니다의 부흥으로 주류 의학 합니다. 예를 들어, 괴로움 인식 면역 기능6 또는7약물 환자의 지를 예측할 수 있습니다. 그럼에도 불구 하 고, 생물 의학 연구원은 크게 맛을 무시 있다. 이 부주의 수 있습니다, 부분적으로,이 감각 시스템 이해에 이른 진행 실험 심리학8, 필드는 의학에 그 수 있습니다 상대적으로 익숙한 뿌리가 있다는 사실을 반영 했다. 또한, 맛에 새롭게 관심 있다 되죠 이전 방법10, 구축 표준화 된 맛 방법9 종합 동안 길고 임상 설정에 대 한 부적 절 한 있습니다. 마지막으로, 맛 조치에 신뢰 때문에 과목 그들의 자신의 경험에 대 한 보고서 및 그들의 관측의 유효성 검사는 지금까지 부족 했던 약한 될 수 있습니다. 우리의 희망은 조사 또는 임상 쉽게 관리할 수 있는 간단한 측정 두 성분과 인기에서 얻게 될 것 이다. 여기 세 부분에는 간단한 맛 시험 프로토콜 설명: 구강, 맛 테스트 및 유효성 검사 단계 선천적인된 유전자 형을 사용 하 여 평가. 첫째, 우리는 의학, 감각 실험 심리학 및 유전자 형 및 유전학을 사용 하 여 응답의 유효성 검사에서 간단한 사례를 병합이 절차에 대 한 생물 학적 컨텍스트를 제공 합니다.
효과적인 맛 시험 명백한 구두 질병, 빨갛게, 붓기와 변색에 대 한 간략 한 임상 평가 포함 해야 그래서 맛 지 각은 입에서 시작 합니다. 7 하위 사이트를 포함 하는 구강: gingiva, 혀, 입, 볼 점 막, 입술 점 막, 하드 미각, 그리고 retromolar trigone의 바닥. 건강 한 참가자 또는 그 잘 정의 된 질병에 초점을 맞춘 인간 맛의 과거 연구 하지만 맛 테스트 루틴에서 의료 시험 되 고, 그것은 절차의 일환으로 구강의 상태를 기록 하는 것이 중요.
혀 자체는 근육 구조 점 막;에 쌌 다 그것의 등 쪽 표면 dotting 용의자, 혀를 그것의 독특한 텍스처 주고 맛 수용 체 세포를 포함 하는 작은 제기 구조입니다. 우리는 그들의 모양에 의해 용의자를 분류: fungiform, filiform, 잎, 그리고 circumvallate. Fungiform 용의자 (FP)는 혀에 있는 anterolaterally 이며 버섯 모양11라운드. 조사자는 FP를 계량 여러 가지 유용한 방법을 출판 그리고 우리 독자 측정 프로토콜12,13,14,,1516에 대 한 이러한 소스를 직접. (Folia), 책의 페이지 측면 후부에 있는 같은 모양의 잎 papillae 혀 표면11. Circumvallate papillae 혀 기지의 고 랑 terminalis에 대형 돔 모양의 구조 점 막 벽 (라틴어 circum, "서라운드," + vallum, "벽")11에 의해 둘러싸여 있습니다. 대부분 수많은 용의자, filiform, 길고 얇은 이며 맛 수용 체를 포함 하지 않습니다.
사람들이 다 혀 해부학. 이 해 부 변이의 소스 알려지지 않은, 하는 동안 결정 됩니다 일부 선천적인된 유전자 변이 의해 조사 중 장신 쌍둥이17일 쌍둥이 60% 일치 중 혀 해부학의 용어 색인 31%를 보고와. 적어도 하나의 유전자 질환 (가족 dysautonomia) 맛 용의자18,,1920의 선 천 성 결핍 귀착되는 희소 한, 및 젖꼭지 밀도 또한 사람들, 다릅니다. 따라서, psychophysical 테스트를 수행 하기 전에 간단한 평가의 일환으로 FP의 존재를 확인 하 고 상대적 크기 및 혀의 색깔과 구강 질환의 증거를 도움이 됩니다.
맛 용의자는 감각 포함 시작 맛 감각을 자극 하는 때 세포. 인간은 맛의 적어도 5 개의 클래스를 감지 할 수 있다: 짠 맛, 신 맛, 쓴, 달콤한, 및 umami. 반면 짠 맛, 달콤한, 그리고 umami 맛 염화 나트륨, 포도 당, 아미노산, 포함 된 귀중 한 음식 근원의 존재를 신호 각각, 쓰라 림과 까다로움 신호 잠재적인 독 소 및 산의 존재의 세균성 분해에서 음식, 각각, 혐오 행동21유도. 짠 맛과 신 맛 맛 세포의 어떤 종류에서 발견 하는 이온 채널의 활성화를 통해 불리고는, 내가 뿐만22,23세포는 소금 변환의 이해 진화 하 고 형식이 필요할 수 있습니다. 쓴, 달콤한, 그리고 umami는 특정 취향에 맞춘 각 타입 II 맛 세포에 G 단백질 결합 된 수용 체의 활성화에서 발생 한다. 3 특정 수용 체의 소 단위의 Heterodimers transduce 정하고 umami 쓴 화합물 25 다른 쓴 수용 체24의 그룹을 활성화 하는 동안. 이 쓴 수용 체는 여러 쓴 화합물에 응답할 수 있습니다 그리고 단일 쓴 화합물은 종종 하나 이상의 수용 체25자극. 맛의 분자 기초에 대 한 지식의 최근 확장에도 불구 하 고 소설 경로26 및 전통적인 5 맛 특성 (예를 들어, 칼슘27 또는 지방산28 지 각)을 넘어 새로운 발견 수 있습니다 앞에 거짓말.
수용 체의 맛 가족의 적어도 두 개의 놀라운 측면이 있다: 이러한 수용 체 수 DNA 순서에서 현저 하 게 다른 사람들, 그러므로 작동 그리고 신체의 많은 조직 표현 이러한 유전자21,29 를 코드 하는 유전자 , 30 , 31. 이러한 extraoral 사이트 포함 두뇌, 갑 상선, 위와 더 낮은 호흡 기관, 그리고 많은 다른 사람의 사이에서 위장21,29,,3031. 동안이 위치에서 맛 수용 체는 전통적인 의미에서 맛 인식에 참여 하지 않는, 그들은 아마 현지 화학 환경29,32를 감지 한다. 상부 호흡기의 ciliated 상피 쓴 수용 체 T2R38을 표현 하는 예를 들어 (쓴 맛 수용 체 38)는 박테리아에 의해 생성 하는 화합물을 응답 하 고 같이 타고 난 면역 반응32영향 mucociliary 클리어런스 및 항균 펩 티 드 및 질소 산화물의 수준을 증가. 이 만성 rhinosinusitis에 대 한 의료 의미, 만성 세균성 감염 질환 및 상부 호흡기와 paranasal sinuses의 염증이 있습니다.
맛에 특정 한 관련성의 시험 여기 설명 T2R38 쓴 맛 수용 체, TAS2R38 유전자에 의해, 유전 가변성 및 따라서 변수 맛 감도 전시. 쓴 화합물 phenylthiocarbamide (PTC)에 대 한 지 각 차이 화학자 아서 폭스33;에 의해 처음으로 설명 했다 나중이 화합물34T2R38 수용 체 주 작동 근으로 확인 되었습니다. 개인 차는 3 단 하나 뉴클레오티드 동 질 다 상, 각 항복 아미노산 대체 (A49P, A262V, 및 I296V; TAS2R38 유전자의 DNA 순서에서 발생 A: 알라닌, p: 프롤린, v: Valine, i: 이소류신). 두 가지 일반적인 haplotypes 결과, PAV 및 AVI, PAV/PAV 개인 PTC ("감별사")에 매우 민감한 되 고, AVI/AVI 개인 되 고 상대적으로 구분 ("비-시음"), 및 그들의 감도에서 더 많은 변수를 되 heterozygous AVI/PAV 개인 35. 쓰라린 인식, 예, 맛 수용 체 TAS2R19 유전자에 의해 T2R19에 영향을 미치는 유전 변이의 더 많은 예제 마찬가지로 전시 유전 가변성 및 쓴 화합물으로 다른 맛 감도 말라리아36. 마찬가지로, TAS2R31 에서 높은 힘 감미료37,,3839중 하나의 인식된 괴로움을 영향을 줍니다.
여기는 환자의 임상 의학, 실험 심리학, 유전학에서 높은 수율 프로토콜에 그리는 맛의 감각을 특성화 하는 빠른 방법에 설명 합니다.

<table border="0" cellpadding="0" cellspacing="0" width="850">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Disposable diagnostic penlight</td>
			<td style="width:163px;">Primacare</td>
			<td style="width:124px;">DL-9223</td>
			<td style="width:259px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">UltraLite Pro headlight</td>
			<td style="width:163px;">Integra LifeSciences</td>
			<td>AX2100BIF</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Millipore Q-Gard 2 water purification system</td>
			<td>EMB Millipore</td>
			<td style="width:124px;">QGARD00D2</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Denatonium benzoate&#160;</td>
			<td>Sigma Aldrich</td>
			<td>D5765</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Phenylthiocarbamide</td>
			<td>Sigma Aldrich</td>
			<td>P7629</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Quinine hydrochloride dihydrate</td>
			<td>Sigma Aldrich</td>
			<td>Q1125</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sodium Chloride</td>
			<td>Sigma Aldrich</td>
			<td>S1679</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:307px;">Sucrose</td>
			<td>Sigma Aldrich</td>
			<td>S0389</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Glass scintillation vials</td>
			<td>Thomas Scientific</td>
			<td>1230L59</td>
			<td>Same as Wheaton catalog no. 986580</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Oragene Discover OGR-500 DNA collection kit</td>
			<td>DNA Genotek</td>
			<td style="width:124px;">OGR-500</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">prepIT L2P Protocol reagents</td>
			<td>DNA Genotek</td>
			<td>PT-L2P-5</td>
			<td></td>
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			<td>ThermoFisher Scientific</td>
			<td>C___8876467_10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rs1726866 TaqMan SNP genotyping assay</td>
			<td>ThermoFisher Scientific</td>
			<td>C___9506827_10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rs10246939 TaqMan SNP genotyping assay</td>
			<td>ThermoFisher Scientific</td>
			<td>C___9506826_10</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">rs10772420 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

의학 및 생물 의학 연구, 및 그것의 유전 토대에 대 한 새로운 지식을 맛의 신흥 중요성 고전적인 맛 테스트 방법 두 가지 방법으로 보완 하기 위해 우리 동기는. 첫째, 우리 입, 혀, 맛 용의자 있는지 확인 하 고 관련 질병의 증거를 참고를 포함 하 여의 간략 한 평가 수행 하는 방법을 설명 합니다. 둘째, 우리 인식된 괴로움 강도와 선된 수용 체 genotypes 비교 보고서에서 맛 테스트 데이터의 유효성을 검사 하는 유전학에 그립니다. 데이터 수집 오류, 산만된 과목 또는 이해 하거나 지시 하지 그 유전자 형의 객관적인 측정 및 맛 경험의 주관적인 보고서 간의 부조화 식별할 수 있습니다. 우리의 기대는 정기적으로, 맛을 평가 하는 연구자와 임상 및 유효한 맛 테스트 설득 수 있습니다 맛 테스트 청력 및 시력에 대 한 테스트로 일반적으로. 마지막으로, 맛 수용 체를 표현 하는 신체의 많은 조직, 때문에 맛 응답 본문에 다른 조직 감도 대 한 프록시를 제공 있으며, 급속 한, 포인트-중-케어 테스트 가이드 진단 및 맛을 평가 하는 연구 도구 역할을 함으로써, 단백질 기능입니다.

취향 테스트 결과 평가 모든 과목에 대 한 풀링된 되어 있다 (n = 840)와 유전자 형 분리 후 제시 됩니다. 전체 데이터 세트 https://carayata.shinyapps.io/TasteBoxplots/에서 액세스할 수 하 고 각 tastant 평가 이며 TAS2R38 및 TAS2R19 genotypes 검토할 수 있습니다. 결과 TAS2R38 수용 체 유전자 형 (그림 3)에 의해 그룹화 하는 과목 가운데 PTC에 대 한 지 ?...

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

Taste Exam: A Brief and Validated Test

맛 시험: 짧고 검증 테스트

이 프로토콜 인간 맛 응답을 측정 하 고 간단한 해 부 평가, 짧은 맛 테스트 및 유효성 검사 방법의 보고 된 감각과 맛 수용 체 유전자를 사용 하 여 포함.

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

This method can help answer key questions in the medical and sensory field about each person's sense of taste. The main advantage of this technique is its brevity and suitability for biomedical research. The implications of this technique extend towards the diagnosis and treatment of chronic rhino sinusitis because taste receptors are part of the innate immune system.I will be demonstrating the procedure on our lab member David and Lauren will be making the taste solutions. Begin by instructing the subject to open their mouth. Use a light to illuminate the oral cavity and examine the seven sub-sites of the area.Here the dorsal tongue is easily visible. Using a tongue depressor to depress the tongue the posterior pharynx becomes visible. Next lateralizing the tongue allows exposure of the left retromolar trigone and the left buccal mucosa.Here the anterior gingiva and the labial mucosa are visualized. As the subject's tongue is lifted the ventral tongue surface becomes visible. And this allows for examination of the subjects floor of mouth.The tongue is then lateralized again to examine the right retromolar trigone and the right buccal mucosa. And lastly the hard and soft palate are visualized. Note lesions, abrasions, and masses or signs of inflammation.Use a light source to visualize the dorsal surface of the tongue and identify the presence or absence of fungiform papillae or smooth surface of the tongue. Finally write down the results of oral cavity examination before proceeding with taste testing. Begin by preparing the taste solutions using a volumetric flask for each to ensure precision of concentrations.Dissolve samples using ultra pure water. Wrap any light sensitive solutions with foil to reduce exposure to light. Then store the taste solutions at four degrees Celsius.To identify common errors in solution preparation fill one tasting cup with the old solution and one with the new solution. Taste each solution to verify they are identical in strength. Next, label vial caps with a circular sticker according to the order of presentation and a color code.In order to match the vial labels with the labels on the taste questionnaire. Package samples by placing them into their respective carriers. Next prepare the taste questionnaire using a category scale for rating taste intensity and a forced choice for identifying the taste quality of each tastant.Lastly, place circular labels of the appropriate color and number next to the appropriate sample in the taste questionnaire. Provide the subject with the sample box, a bottle of water, an empty cup, pen, and taste questionnaire containing entries for 12 samples. Explain the testing procedure and inform the subject that they will be asked to rate both the intensity and quality of each tastant.Let them know that they may not experience all qualities. Start the test by asking the subject to rinse their mouth twice with water and spit it out in the cup provided. Have them pour all of sample one into their mouth and hold it there for five seconds before spitting the solution into the cup.Tell them not to gargle or swallow the solution. Then instruct the subject to circle one of the 13 vertical lines corresponding with the sample's intensity. On a scale of 0 to 12.From no intensity at all to extremely intense. And choose a single quality to describe the taste. Next ask the subject to rinse their mouth with water twice before proceeding to the next sample.Observe the subject tasting and rating sample one of water. Should the rating deviate from no intensity at all and no flavor, reiterate the questionnaire instructions before allowing the test to proceed. After all samples have been tasted review the finished questionnaire for completeness.Finally, score intensity ratings on a scale of 0 to 12 from the vertical lines that the subject circled. And average the two intensity ratings for each tastant for later analysis. Results of the taste test confirm the existence of perceptual taste differences for the bitter compound, Phenylthiocarbamide or PTC among subjects grouped by the TAS2R38 receptor genotype.Ratings of PTC intensity are significantly different across three genotypes. The motivation behind this study was to try to assess or to develop a rapid point of service exam that we could do in the clinic that would assess the function of this receptor that we believe plays a role in the development of chronic sinusitis. The idea here is could we use this taste test to predict which of our patients are at high risk for developing sinusitis and more importantly, which patients are good surgical candidates for us to do our sinus surgery on, or which ones will do poorly with our sinus surgery.So we're trying to use this taste test to prognosticate patient's sinal-nasal disease. Once that we went through all of the taste testing we confirmed that by genotyping the patients. And as you can see in figure three for the most part the patient's genotype follows the pattern of their phenotype.Meaning the more bitter they tasted they tended to be dominant for this receptor. However, there are some outliers. Showing that there are functional homozygotes, PAV/PAVs that said that they didn't taste PTC and on the flip side of that there were some nonfunctional AVI/AVI individuals that tasted PTC with bitterness.So that means that whatever taste test we do develop if there are patients that are not fulfilling our expectations that we probably wanna confirm the taste test by genotyping them for T2R38. Once mastered the exam and taste test can be performed in under 30 minutes. After watching this video you should have an understanding of how to perform an oral exam, note evidence of disease and perform a brief taste test.

Research

JoVE Journal

Medicine

Making Sense of Listening: The IMAP Test Battery

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the abilities to extract and use the information encoded in the auditory signal. We refer to these as listening skills (or auditory processing AP). Deficits in these skills are associated with delayed language and literacy development, though the nature of the relevant deficits and their causal connection with these delays is hotly debated.
When a child is referred to a health professional with normal hearing and unexplained difficulties in listening, or associated delays in language or literacy development, they should ideally be assessed with a combination of psychoacoustic (AP) tests, suitable for children and for use in a clinic, together with cognitive tests to measure attention, working memory, IQ, and language skills. Such a detailed examination needs to be relatively short and within the technical capability of any suitably qualified professional. Current tests for the presence of AP deficits tend to be poorly constructed and inadequately validated within the normal population. They have little or no reference to the presenting symptoms of the child, and typically include a linguistic component. Poor performance may thus reflect problems with language rather than with AP. To assist in the assessment of children with listening difficulties, pediatric audiologists need a single, standardized child-appropriate test battery based on the use of language-free stimuli.
We present the IMAP test battery which was developed at the MRC Institute of Hearing Research to supplement tests currently used to investigate cases of suspected AP deficits. IMAP assesses a range of relevant auditory and cognitive skills and takes about one hour to complete. It has been standardized in 1500 normally-hearing children from across the UK, aged 6-11 years. Since its development, it has been successfully used in a number of large scale studies both in the UK and the USA. IMAP provides measures for separating out sensory from cognitive contributions to hearing. It further limits confounds due to procedural effects by presenting tests in a child-friendly game-format. Stimulus-generation, management of test protocols and control of test presentation is mediated by the IHR-STAR software platform. This provides a standardized methodology for a range of applications and ensures replicable procedures across testers. IHR-STAR provides a flexible, user-programmable environment that currently has additional applications for hearing screening, mapping cochlear implant electrodes, and academic research or teaching.

A test battery (IMAP) for performing an in-depth assessment of auditory and cognitive abilities contributing to listening skills is described. It is quick to administer, child-friendly and free from linguistic confounds. Stimulus generation and protocol management are controlled via a software platform (IHR-STAR) to ensure replicable procedures.

IMAP 테스트 배터리 : 듣기의 감각을 만들기

The ability to hear is only the first step towards making sense of the range of information contained in an auditory signal. Of equal importance are the ...

연구

의학

Technique to Collect Fungiform (Taste) Papillae from Human Tongue

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic processes that prepare the alimentary canal for digestion. Steady progress is being made towards understanding the early biochemical and molecular events underlying taste transduction (for a review, Breslin and Spector, 20081). However, progress to date has largely resulted from animal models. Yet, since marked differences in receptor specificity and receptor density vary among species, human taste transduction will only be understood by using human taste tissue. Here we describe a biopsy technique to collect human fungiform papillae, visible as rounded pink anterior structures, about 0.5 mm in diameter that contain taste buds. These biopsied papillae are used for several purposes including the isolation of viable taste bud cells, in situ hybridization, immunohistochemistry and, through techniques of molecular biology, the identification of taste-specific novel proteins.

Technique to Collect Fungiform Taste Papillae from Human Tongue

Knowledge of molecular mechanisms underlying gustatory transduction has recently enjoyed significant advances, largely due to using animal models. However, the wide diversity in taste sensitivity and specificity among mammals warrants studies in human tissue. We describe a biopsy technique to collect living taste cells from the papillae on human tongue.

인간 혀에서 Fungiform (맛) 용의자를 수집하는 기술

The sense of taste is critical for human life. It informs the body about the quality of food that will be potentially ingested and stimulates metabolic ...

The Trier Social Stress Test Protocol for Inducing Psychological Stress

This article demonstrates a psychological stress protocol for use in a laboratory setting. Protocols that allow researchers to study the biological pathways of the stress response in health and disease are fundamental to the progress of research in stress and anxiety.1 Although numerous protocols exist for inducing stress response in the laboratory, many neglect to provide a naturalistic context or to incorporate aspects of social and psychological stress. Of psychological stress protocols, meta-analysis suggests that the Trier Social Stress Test (TSST) is the most useful and appropriate standardized protocol for studies of stress hormone reactivity.2 In the original description of the TSST, researchers sought to design and evaluate a procedure capable of inducing a reliable stress response in the majority of healthy volunteers.3 These researchers found elevations in heart rate, blood pressure and several endocrine stress markers in response to the TSST (a psychological stressor) compared to a saline injection (a physical stressor).3 Although the TSST has been modified to meet the needs of various research groups, it generally consists of a waiting period upon arrival, anticipatory speech preparation, speech performance, and verbal arithmetic performance periods, followed by one or more recovery periods. The TSST requires participants to prepare and deliver a speech, and verbally respond to a challenging arithmetic problem in the presence of a socially evaluative audience.3 Social evaluation and uncontrollability have been identified as key components of stress induction by the TSST.4 In use for over a decade, the goal of the TSST is to systematically induce a stress response in order to measure differences in reactivity, anxiety and activation of the hypothalamic-pituitary-adrenal (HPA) or sympathetic-adrenal-medullary (SAM) axis during the task.1 Researchers generally assess changes in self-reported anxiety, physiological measures (e.g. heart rate), and/or neuroendocrine indices (e.g. the stress hormone cortisol) in response to the TSST. Many investigators have adopted salivary sampling for stress markers such as cortisol and alpha-amylase (a marker of autonomic nervous system activation) as an alternative to blood sampling to reduce the confounding stress of blood-collection techniques. In addition to changes experienced by an individual completing the TSST, researchers can compare changes between different treatment groups (e.g. clinical versus healthy control samples) or the effectiveness of stress-reducing interventions.1

This article describes a protocol for inducing psychological stress in participants, which enables researchers to measure psychological, physiological and neuroendocrine responses to stress within single participants or between groups.

심리적 스트레스를 유발을위한 모금 사회 스트레스 테스트 프로토콜

This article demonstrates a psychological stress protocol for use in a laboratory setting.  Protocols that allow researchers to study the biological ...

The &alpha;-test: Rapid Cell-free CD4 Enumeration Using Whole Saliva

There is an urgent need for affordable CD4 enumeration to monitor HIV disease. CD4 enumeration is out of reach in resource-limited regions due to the time and temperature restrictions, technical sophistication, and cost of reagents, in particular monoclonal antibodies to measure CD4 on blood cells, the only currently acceptable method. A commonly used cost-saving and time-saving laboratory strategy is to calculate, rather than measure certain blood values. For example, LDL levels are calculated using the measured levels of total cholesterol, HDL, and triglycerides1. Thus, identification of cell-free correlates that directly regulate the number of CD4+ T cells could provide an accurate method for calculating CD4 counts due to the physiological relevance of the correlates.
The number of stem cells that enter blood and are destined to become circulating CD4+ T cells is determined by the chemokine CXCL12 and its receptor CXCR4 due to their influence on locomotion2. The process of stem cell locomotion into blood is additionally regulated by cell surface human leukocyte elastase (HLECS) and the HLECS-reactive active &alpha;1proteinase inhibitor (&alpha;1PI, &alpha;1antitrypsin, SerpinA1)3. In HIV-1 disease, &alpha;1PI is inactivated due to disease processes 4. In the early asymptomatic categories of HIV-1 disease, active &alpha;1PI was found to be below normal in 100% of untreated HIV-1 patients (median=12 &mu;M, and to achieve normal levels during the symptomatic categories4, 5. This pattern has been attributed to immune inactivation, not to insufficient synthesis, proteolytic inactivation, or oxygenation. We observed that in HIV-1 subjects with &gt;220 CD4 cells/&mu;l, CD4 counts were correlated with serum levels of active &alpha;1PI (r2=0.93, p&lt;0.0001, n=26) and inactive &alpha;1PI (r2=0.91, p&lt;0.0001, n=26) 5. Administration of &alpha;1PI to HIV-1 infected and uninfected subjects resulted in dramatic increases in CD4 counts suggesting &alpha;1PI participates in regulating the number of CD4+ T cells in blood 3.
With stimulation, whole saliva contains sufficient serous exudate (plasma containing proteinaceous material that passes through blood vessel walls into saliva) to allow measurement of active &alpha;1PI and the correlation of this measurement is evidence that it is an accurate method for calculating CD4 counts. Briefly, sialogogues such as chewing gum or citric acid stimulate the exudation of serum into whole mouth saliva. After stimulating serum exudation, the activity of serum &alpha;1PI in saliva is measured by its capacity to inhibit elastase activity. Porcine pancreatic elastase (PPE) is a readily available inexpensive source of elastase. PPE binds to &alpha;1PI forming a one-to-one complex that prevents PPE from cleaving its specific substrates, one of which is the colorimetric peptide, succinyl-L-Ala-L-Ala-L-Ala-p-nitroanilide (SA3NA). Incubating saliva with a saturating concentration of PPE for 10 min at room temperature allows the binding of PPE to all the active &alpha;1PI in saliva. The resulting inhibition of PPE by active &alpha;1PI can be measured by adding the PPE substrate SA3NA. (Figure 1). Although CD4 counts are measured in terms of blood volume (CD4 cells/&mu;l), the concentration of &alpha;1PI in saliva is related to the concentration of serum in saliva, not to volume of saliva since volume can vary considerably during the day and person to person6. However, virtually all the protein in saliva is due to serum content, and the protein content of saliva is measurable7. Thus, active &alpha;1PI in saliva is calculated as a ratio to saliva protein content and is termed the &alpha;1PI Index. Results presented herein demonstrate that the &alpha;1PI Index provides an accurate and precise physiologic method for calculating CD4 counts.

The &amp;#945;-test: Rapid Cell-free CD4 Enumeration Using Whole Saliva

A CD4 enumeration method, the &#x03B1;-test, is described which uses whole saliva to provide rapid and accurate CD4 counts. The &alpha;-test costs pennies and eliminates the need for technical training, costly reagents such as monoclonal antibodies, instrumentation, refrigeration, transport of samples, as well as collection and handling of blood.

α-테스트 : 전체 타액을 사용하여 빠른 셀 프리 CD4의 열거

There is an urgent need for affordable CD4 enumeration to monitor HIV disease. CD4 enumeration is out of reach in resource-limited regions due to the time ...

Flexible Colonoscopy in Mice to Evaluate the Severity of Colitis and Colorectal Tumors Using a Validated Endoscopic Scoring System

The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental endoscopy has been highly useful for studies that require repeated assessments in a single laboratory animal, such as those evaluating mechanisms of chronic inflammatory bowel disease and the progression of colorectal cancer. However, the methods used across studies are highly variable. At least three endoscopic scoring systems have been published for murine colitis and published protocols for the assessment of colorectal tumors fail to address the presence of concomitant colonic inflammation. This study develops and validates a reproducible endoscopic scoring system that integrates evaluation of both inflammation and tumors simultaneously. This novel scoring system has three major components: 1) assessment of the extent and severity of colorectal inflammation (based on perianal findings, transparency of the wall, mucosal bleeding, and focal lesions), 2) quantitative recording of tumor lesions (grid map and bar graph), and 3) numerical sorting of clinical cases by their pathological and research relevance based on decimal units with assigned categories of observed lesions and endoscopic complications (decimal identifiers). The video and manuscript presented herein were prepared, following IACUC-approved protocols, to allow investigators to score their own experimental mice using a well-validated and highly reproducible endoscopic methodology, with the system option to differentiate distal from proximal endoscopic colitis (D-PECS).

Over the past few years, new generation endoscopes have emerged as important diagnostic research aids for evaluating murine colitis and colorectal tumors. We present herein a detailed protocol for endoscopic assessment of inflammation and colorectal tumors in mice, as well as a novel scoring system that uses decimal identifiers to document the endoscopic severity of colitis and colorectal tumors.

검증 된 내시경 채점 시스템을 사용하여 대장염과 대장 종양의 위험을 평가하기 위해 마우스의 유연한 대장 내시경 검사

The use of modern endoscopy for research purposes has greatly facilitated our understanding of gastrointestinal pathologies. In particular, experimental ...

The Multiple Sclerosis Performance Test (MSPT): An iPad-Based Disability Assessment Tool

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report a new test, the Multiple Sclerosis Performance Test (MSPT), which represents a new approach to quantifying MS related disability. The MSPT takes advantage of advances in computer technology, information technology, biomechanics, and clinical measurement science. The resulting MSPT represents a computer-based platform for precise, valid measurement of MS severity. Based on, but extending the Multiple Sclerosis Functional Composite (MSFC), the MSPT provides precise, quantitative data on walking speed, balance, manual dexterity, visual function, and cognitive processing speed. The MSPT was tested by 51 MS patients and 49 healthy controls (HC). MSPT scores were highly reproducible, correlated strongly with technician-administered test scores, discriminated MS from HC and severe from mild MS, and correlated with patient reported outcomes. Measures of reliability, sensitivity, and clinical meaning for MSPT scores were favorable compared with technician-based testing. The MSPT is a potentially transformative approach for collecting MS disability outcome data for patient care and research. Because the testing is computer-based, test performance can be analyzed in traditional or novel ways and data can be directly entered into research or clinical databases. The MSPT could be widely disseminated to clinicians in practice settings who are not connected to clinical trial performance sites or who are practicing in rural settings, drastically improving access to clinical trials for clinicians and patients. The MSPT could be adapted to out of clinic settings, like the patient&#8217;s home, thereby providing more meaningful real world data. The MSPT represents a new paradigm for neuroperformance testing. This method could have the same transformative effect on clinical care and research in MS as standardized computer-adapted testing has had in the education field, with clear potential to accelerate progress in clinical care and research.

The Multiple Sclerosis Performance Test MSPT: An iPad-Based Disability Assessment Tool

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report methodologic details on a new test, the Multiple Sclerosis Performance Test (MSPT). This new approach to the objective of quantification of MS related disability provides a computer-based platform for precise, valid measurement of MS severity.

다발성 경화증 성능 테스트 (MSPT) : 아이 패드 기반 장애 평가 도구

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report a new test, the ...

Determining The Electromyographic Fatigue Threshold Following a Single Visit Exercise Test

Theoretically, the electromyographic (EMG) fatigue threshold is the exercise intensity an individual can maintain indefinitely without the need to recruit more motor units which is associated with an increase in the EMG amplitude. Although different protocols have been used to estimate the EMG fatigue threshold they require multiple visits which are impractical for a clinical setting. Here, we present a protocol for estimating the EMG fatigue threshold for cycle ergometry which requires a single visit. This protocol is simple, convenient, and completed within 15-20 min, therefore, has the potential to be translated into a tool that clinicians can use in exercise prescription.

This protocol describes the electromyographic fatigue threshold which demarcates between nonfatiguing and fatiguing exercise workloads. This information could be used to develop a more individualized training program.

단일 방문 연습 시험을 다음과 근전도 피로 임계 값 결정

Theoretically, the electromyographic (EMG) fatigue threshold is the exercise intensity an individual can maintain indefinitely without the need to recruit ...

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

생쥐 경동맥 풍선 손상 모델은 반대로 혈관 리모델링의 치료를 테스트하는

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

Ultrasound Assessment of Endothelial Function: A Technical Guideline of the Flow-mediated Dilation Test

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a pathological condition characterized mainly by a disruption in the balance between vasodilator and vasoconstrictor substances and is proposed to play an important role in the development of atherosclerotic cardiovascular disease. Therefore, a precise evaluation of endothelial function in humans represents an important tool that could help better understand the etiology of multiple cardio-centric pathologies.
Over the past twenty-five years, many methodological approaches have been developed to provide an assessment of endothelial function in humans. Introduced in 1989, the FMD test incorporates a forearm occlusion and subsequent reactive hyperemia that promotes nitric oxide production and vasodilation of the brachial artery. The FMD test is now the most widely utilized, non-invasive, ultrasonic assessment of endothelial function in humans and has been associated with future cardiovascular events.
Although the FMD test could have clinical utility, it is a physiological assessment that has inherited several confounding factors that need to be considered. This article describes a standardized protocol for determining FMD including the recommended methodology to help minimize the physiological and technical issues and improve the precision and reproducibility of the assessment.

The flow mediated dilation (FMD) test is the most commonly utilized, non-invasive, ultrasound assessment of endothelial function in humans. Although the FMD test has been related with the prediction of future cardiovascular disease and events, it is a physiological assessment with many inherent confounding factors that need to be considered.

내피 기능의 초음파 평가 : 흐름 중재 팽창 시험의 기술 가이드 라인

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a ...

A Blood-based Test for the Detection of ROS1 and RET Fusion Transcripts from Circulating Ribonucleic Acid Using Digital Polymerase Chain Reaction

We have developed novel methods for the isolation and characterization of tumor-derived circulating ribonucleic acid (cRNA) for blood-based liquid biopsy. Robust detection of cRNA recovered from blood represents a solution to a critical unmet need in clinical diagnostics. The test begins with the collection of whole blood into blood collection tubes containing preservatives that stabilize cRNA. Cell-free, exosomal, and platelet-associated RNA is isolated from plasma in this test system. The cRNA is reverse transcribed to complementary DNA (cDNA) and amplified using digital polymerase chain reaction (dPCR). Samples are evaluated for both the target biomarker as well as a control gene. Test validation included limit of detection, accuracy, and robustness studies with analytic samples. The method developed as a result of these studies reproducibly detect multiple fusion variants for ROS1 (C-Ros proto-oncogene 1; 8 variants) and RET (rearranged during transfection proto-oncogene; 8 variants). The sample processing workflow has been optimized so that test results can consistently be generated within 72 hours of sample receipt.

Detection of circulating ribonucleic acid (cRNA) from blood is an unmet need in clinical diagnostics. Here we describe methods that characterize cRNA from non-small cell lung cancer patients using sensitive and specific digital polymerase chain reaction. The tests meet design requirements to detect fusion variants within 72 hours.

혈액 기반 ROS1의 검출을 위한 테스트 및 RET Ribonucleic 산 디지털 연쇄 반응을 사용 하 여 순환에서 퓨전 성적표

We have developed novel methods for the isolation and characterization of tumor-derived circulating ribonucleic acid (cRNA) for blood-based liquid biopsy. ...