저자는 스캔 소프트웨어를 개선와의 지속적인 지원을위한 마스터 참조 모델 및 Alicona 회사 생산을위한 치과 기술자 니콜라 Lanfranconi 감사합니다.

1. 표본 준비 / 자료 <ol><li> 시편 주위에 편평한 기초를 적용합니다. 스캔 테이블에 시료를 놓습니다. 수평면 (그림 1)에 교합면 배향. </li></ol> 이. 소프트웨어 분석 <ol><li> 프로그램을 시작하고 실험실 측정 모듈 (그림 2) </li><li> 주사 테이블의 중앙에 시편을 위치. </li><li> 올바른 배율의 렌즈를 선택합니다. 큰 개체의 경우, 전체 아치 스캔처럼 5 배 목표를 사용합니다. </li><li> 시편의 표면이 라이브 뷰 창 (그림 3)에 표시 될 때까지 3D 마우스를 사용하여 스캔 광학 이동합니다. </li><li> 최적의 검색 매개 변수를 달성하기 위해 노출 및 명암을 설정하는 센서 컨트롤을 사용합니다. 금속 표면의 경우, 400 및 800 마이크로 초 0.3과 0.8 사이의 대조 사이의 노출 (그림 4)를 사용합니다. </li><li> 버튼을 선택하여 이미지 품질을 점검&quot;실시간 미리보기 이미지 표시 품질&quot;. </li><li> 는 &#39;측정 제어 &quot;올바른 매개 변수를 설정합니다. 측정 방식은 &quot;3D 데이터 집합&quot;이고; 하여 ImageField 유형은 &quot;일반하여 ImageField&quot;(그림 5)입니다. </li><li> &quot;새로운하여 ImageField&quot;를 클릭하십시오. </li><li> 스캔 볼륨을 정의합니다. 이 소프트웨어는 측정 볼륨의 경계 좌표를 필요로한다. <ol><li> 최고와 최저 스캔 수준에 표본을 이동하고 두 지점에서 &quot;추가 위치&quot;를 클릭합니다. &quot;Z 범위&quot;값은 스캔 볼륨의 실제 높이 (그림 6)를 보여줍니다. </li><li> 스캔 볼륨의 XY boundings에 표본을 이동합니다. X-축 및 Y-축 길이를 정의하는 &quot;추가 위치&quot;를 클릭하세요. 주사 부피의 사이즈는 &quot;정보&quot;탭에 도시되고 X-축 및 Y-축에서 1cm 의해 시험편의 치수를 초과한다. </li></ol></li><li> &quot;포인트&quot;번호를 확인합니다. 이 소프트웨어는 하나 &quot;하여 ImageField&quot;스캔 스캔 억 표면 점 할 수있다. 실제 수점으로이 제한을 초과합니다. 포인트 크기의 &quot;측면 다운 샘플링&quot;에 의해 측정 지점의 수를 &quot;죽이다&quot;. <ol><li> (그림 7) &quot;고급 설정&quot;을 클릭합니다. </li><li> &quot;포인트&quot;번호는 (100) M (그림 7) 이하로 감소 될 때까지 오른쪽에있는 &quot;옆 다운 샘플링&quot;슬라이더를 이동합니다. 다운 샘플링은 검사 개체의 더 큰 픽셀 크기의 결과로, 표면 지점의 측 방향 해상도를 감소시킨다. 최적의 검사 결과를 보장하기 위해, 수직 해상도가 증가한다. </li></ol></li><li> &quot;시작 측정&quot;을 클릭합니다. 이것은 &quot;미리보기 모드&quot;를 시작합니다. 소프트웨어는 선택된 X-및 Y-축 치수 프리 스캔을 수행한다. </li><li> 프리 스캔이 완료되면, 관심있는 지역을 선택합니다. 이 파일 크기와 스캔 시간 (그림 8)를 감소하는 데 도움이됩니다. </li><li> 편평한 기초를 포함하는 시료 주위의 표본과 두 개의 측정 분야, (그림 9)를 제외하고는 사전 스캔의 모든 부분을 선택합니다. </li><li> 스캔을 시작하려면 &quot;시작&quot;을 클릭합니다. </li><li> 마우스를 사용하여 &quot;ImageViewer&quot;의 검사를 제어하고 마우스 왼쪽 버튼 (그림 10)를 누릅니다. </li><li> 창을 닫고 &quot;만보기 의사 색상&quot;을 클릭합니다. </li><li> &quot;설정&quot;과 &quot;의사 색칠하기&quot;를 클릭하고 &quot;반복&quot;을 선택하십시오. </li><li> &quot;최대.&quot;로 설정 0.2로 설정하고 값을 &quot;적용 범위&quot;를 클릭합니다. </li><li> 반복을 제어, 같은 경사와 소재 지역과 동일해야합니다. 특히, 시편 주변의베이스는 균일 한 반복 (그림 11)을 표시해야합니다. </li><li> &quot;데이터베이스&quot;를 클릭하고 해당 폴더로 스캔을 저장합니다. </li><li> 필요한 경우, 다른 파일 형식으로 스캔을 보냅니다. 를 클릭하십시오 &quot;File/Export/3D 데이터로 / ....&quot; 프로토콜은 그 시점에서 일시 이상 계속 될 수있다. </li></ol> 3. 차이 분석 <ol><li> 기본 버려야하는 &quot;3D 편집기&quot;를 사용합니다. 이 영역의 차이를 사용하지 않을분석. </li><li> 비교 및 분석하는 두 가지 검사를, &quot;차이 측정&quot;소프트웨어 (그림 12)를 시작합니다. </li><li> 비교하는 두 번째 모델을 선택합니다. </li><li> 모델의 첫 경기를 수행하기 위해 &quot;자동 거친 정렬&quot;을 클릭합니다. </li><li> &quot;수동 조정&quot;을 클릭하고이 같은 방향 (그림 13)가 될 때까지 회전 및 이동로 모델을 맞 춥니 다. </li><li> &quot;자동 정렬&quot;을 클릭하고 최적의 모형 일치에 가장 적합한 알고리즘을 시작하려면 &quot;적용&quot;. </li><li> 두 일치하는 모델의 차이의지도를 볼 수 &quot;차이점&quot;을 클릭합니다. </li><li> (그림 14)의 편차를 표시하는 적절한 색상 범위를 선택합니다. 분석을위한 스크린 샷과 같은 시각적 편차 패턴을 저장합니다. </li></ol> 차이의 통계 값을 표시하는 &quot;통계&quot;를 클릭하십시오. 1 μM 클래스 크기 선택과 통계적 비교를 위해 텍스트 파일 히스토그램 데이터를 저장 (도 15).

<ol>
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저자는 더 경쟁 재정적 이익 또는 그 밖의 충돌이 없음을 선언합니다.

정확도는 치과 의학의 기본 요구이다. 참고 스캐너는 높은 진실성과 정확성에 크고 작은 물체를 스캔 할 수있다. 최적의 주사 방법으로, 심지어 개별 형태학 상세한 치아 표면은 높은 해상도 및 반복성으로 스캔 할 수있다. 스캐너의 상이한 배율 수준, 매크로 및 마이크로 구조 형태를 취득 할 수있다. 이 모델 다양한 재료를 검사하는 것이 가능하다. 참고 스캐너의 정확도 평...

정확도는 치과 의학의 많은 분야에 큰 관심입니다. 치아 경조직을 대체하는 것은 적절한 기능을 보장하고 더 남아있는 치아 구조 1,2를 파괴 방지하기 위해 정확한 피팅 의지가 필요합니다. 고정 부분 틀니와 전체 보철물 정확한 준비된 치아 또는 임플란트 3와 같은지지 구조에 맞는 경우 특히 중요합니다. 매우 정확한 재생 특히 치과 노출 수와 치과 실험실 워크 플로의 필드에, 필요한 이유입니다. 그러나 치과 치료의 다른 분야는 치료의 성공을 확인하고 새로운 치료 전략, 예를 들어, 소프트 및 하드 조직 확대, 부식 및 마모 모니터링, 치주 치료 및 교정 치료 4,5을 평가하기 위해, 진실하고 정확한 운율 결과에서 혜택을 누릴 수 있습니다. 이 분야의 많은에서는, 현재의 검증 절차는 캘리퍼스 또는 현미경 6,7와 직선 거리를 측정합니다. 이 메타ODS는 몇 측정점과 테스트 영역의 3 차원 (3D)의 변화의 제한된 정보에 한정된다. 새로운 측정 방법은 테스트 개체의 8,9의 전체 표면의 광 또는 방사선 캡처 있습니다. 여기서, 전체면 또는 양을 측정하고 컴퓨터 화면에 3D 객체로 표시된다. 선형 측정이 가능하다뿐만 아니라 다른 검사 시간에서 모델 superimpositions. 이 중첩와 함께, 모든 검사 점에서 표면의 변화에​​ 대한 평가가 가능합니다. 이것은 특정 지역을 감시 또는 모든 세 축 좌표에서 변형 표시의 가능성을 가능하게한다. 또한 체적 변화가 열을 측정 할 수있다. 이러한 새로운 방법으로 제한하는 점은 시험 물체를 캡처하는 데 사용되는 스캐너의 정확성이다. 기준 스캐너의 정확도 내에서 변화 없음 시험 물체 또는 스캔 에러의 변화로 분할 될 수 없다. 스캔 정확도는 종종 마뉘에 의해 주어진 값이다선급 검사원은 작은, 보정 오브젝트 (11)를 스캔에서 파생. 치열 궁 같은 큰 물체를 스캔 할 때이 최소한의 검사 오류가 다릅니다. 정확도는 진실성과 정확성으로 구성되어 있습니다. 진도는 실제 형상에서 스캔 한 개체의 편차이다. 정밀 반복 검사 (ISO 5725-1) 사이의 편차이다. 본 연구에서는, 포커스 변화 스캐닝 기술을 기반으로하는 새 광학 기준 스캐너, 높은 정확도로 전체 아치 모델에 하나의 치아에서 표본을 검사하기 위해 도입되었다. 이 기준 스캐너는 기존의 디지털 기술 12-14에서 치과 교합과 치과 재료의 마모와 관련된 실제 프로젝트를위한 치과 인상의 정확성을 비교, 여러 연구를위한 자료로 사용되었다. 본 연구의 목적은 치과 연구 분야에서이 장치를 사용하는 기본적인 기준 스캐너의 정밀도 정보와 일부 가능성을 제공하는 것이었다.

<table border="0" cellpadding="0" cellspacing="0" width="669">
	<tbody>
		<tr>
			<td>Reference model</td>
			<td></td>
			<td></td>
			<td>individual non-precious metal model, derived from a patient impression</td>
		</tr>
		<tr>
			<td>Araldit repair</td>
			<td>Huntsmen Advanced Material, Basel, Switzerland</td>
			<td></td>
			<td>used for making the base of the reference model</td>
		</tr>
		<tr>
			<td>CamBase</td>
			<td>Dentona, Dortmund, Germany</td>
			<td></td>
			<td>Type IV dental ston for pouring conventional impressions</td>
		</tr>
		<tr>
			<td>Identium</td>
			<td>Kettenbach, Eschenburg, Germany</td>
			<td></td>
			<td>Vinylsiloxanether impression material for conventional impression</td>
		</tr>
		<tr>
			<td>inEOS model holder</td>
			<td>Sirona Dental Systems, Bensheim, Germany</td>
			<td></td>
			<td>used for fixing stone models at the reference scanner</td>
		</tr>
		<tr>
			<td>Accutrans</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>used for making the base of thestone models</td>
		</tr>
		<tr>
			<td>President putty</td>
			<td>Coltene Whaledent, Altst&#228;tten, Switzerland</td>
			<td></td>
			<td>mix with accutrans for betterstability of the base</td>
		</tr>
		<tr>
			<td>Alicona Infinite Focus</td>
			<td>Alicona Imaging, Graz, Austria</td>
			<td></td>
			<td>Reference scanner&#160;</td>
		</tr>
	</tbody>
</table>

accuracy in dental medicine, a new way to measure trueness and precision

참고 스캐너는 절차의 많은을 확인 치과 의학에 사용됩니다. 주요 관심은 치과 보철물의 기본 역할로 인상 방법을 확인하는 것입니다. 많은 기준 스캐너의 전류 제한은 전체 치과 아치 또는 상세한 치아 표면을 평가하기 위해 제한된 가능성 같은 정확도 스캐닝 큰 개체의 부족이다. 초점의 변화 스캐닝 기술을 기반으로하는 새로운 기준 스캐너, 가장 높은 지역 및 일반 정확성과 관련하여 평가 하였다. 특정 검사 프로토콜은 치과 노출에서 원래의 치아 표면을 스캔 테스트되었습니다. 또한, 다른 모델 물질이 확인되었다. 결과는 평균 진도 5.3 ± 1.1 μM의 편차 및 전체 아치 스캔의 경우 정밀도를 1.6 ± 0.6 μM의 참조 스캐너의 높은 검색 정확도를 보여 주었다. t 현재 치과 인상 방법은 훨씬 높은 편차 (12.5 ± 2.5 μm의 : 20.4 ± 2.2 μM, 정밀 진위를) 보여 주었다한 기준 스캐너의 내부 검사의 정확도. 단일 치아 표면 같은 작은 물체가 미란 성 및 연마 치아 표면 손실을 평가하는 시스템을 가능하게 더 높은 정밀도로 검사 할 수있다. 기준 스캐너 치과 연구 분야의 많은 차이를 측정 할 수있다. 높은 지역과 일반 정확도와 결합 된 다른 배율 수준은 전체 아치 변화까지 단일 치아 또는 보철물의 변화를 평가하기 위해 사용될 수있다.

그림 16A는 석고 시료의 검사를 보여줍니다. 최적의 검색 매개 변수를 확인하기 위해, 재료의 반복을 제어 할 수 있습니다. 시편의 섹션은 서로 다른 밝기와 대비 설정을 스캔 및 반복성은 최적의 스캔 설정을 찾기 위해 각 스캔 후 확인됩니다. 금속 표면은 높은 콘트라스트 대비 낮은 1.0 석고 또는 수지 재료와 최상의 결과를 보여줍니다. 그림 16b는 시험 검사 후 최적의 검...

Watch this Scientific Journal Video about Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision at JoVE.com

Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

정확도는 치과 의학의 중요한 요구이다. 정확성을 확인하기 위해, 참조 스캐너가 필요합니다. 이 문서에서는 높은 진도​​와 정밀도를 가진 치과 형태학의 광범위한 다양성을 얻기위한 조정 스캔 방법을 사용하여 새 기준 스캐너를 제공한다.

진도 및 정밀 측정하는 치과, 새로운 방법의 정확도

Reference scanners are used in dental medicine to verify a lot of procedures. The main interest is to verify impression methods as they serve as a base ...

accuracy-dental-medicine-new-way-to-measure-trueness

Research

JoVE Journal

Medicine

13.3K 조회수.  University of Zürich.  정확도는 치과 의학의 중요한 요구이다. 정확성을 확인하기 위해, 참조 스캐너가 필요합니다. 이 문서에서는 높은 진도​​와 정밀도를 가진 치과 형태학의 광범위한 다양성을 얻기위한 조정 스캔 방법을 사용하여 새 기준 스캐너를 제공한다. 

비디오: Accuracy in Dental Medicine, A New Way to Measure Trueness and Precision

Method to Measure Tone of Axial and Proximal Muscle

The control of tonic muscular activity remains poorly understood. While abnormal tone is commonly assessed clinically by measuring the passive resistance of relaxed limbs1, no systems are available to study tonic muscle control in a natural, active state of antigravity support. We have developed a device (Twister) to study tonic regulation of axial and proximal muscles during active postural maintenance (i.e. postural tone). Twister rotates axial body regions relative to each other about the vertical axis during stance, so as to twist the neck, trunk or hip regions. This twisting imposes length changes on axial muscles without changing the body's relationship to gravity. Because Twister does not provide postural support, tone must be regulated to counteract gravitational torques. We quantify this tonic regulation by the restive torque to twisting, which reflects the state of all muscles undergoing length changes, as well as by electromyography of relevant muscles. Because tone is characterized by long-lasting low-level muscle activity, tonic control is studied with slow movements that produce "tonic" changes in muscle length, without evoking fast "phasic" responses. Twister can be reconfigured to study various aspects of muscle tone, such as co-contraction, tonic modulation to postural changes, tonic interactions across body segments, as well as perceptual thresholds to slow axial rotation. Twister can also be used to provide a quantitative measurement of the effects of disease on axial and proximal postural tone and assess the efficacy of intervention.

We have developed a device (Twister) to study the regulation of tonic muscle activity during active postural maintenance. Twister measures torsional resistance and muscular responses in standing subjects during twisting of the body axis. The device can be flexibly configured to study various aspects of tonic control across the neck, trunk, and/or hips.

축방향 및 근위 근육 톤을 측정하는 방법

The control of tonic muscular activity remains poorly understood.  While abnormal tone is commonly assessed clinically by measuring the passive resistance ...

연구

의학

Screening for Melanoma Modifiers using a Zebrafish Autochthonous Tumor Model

Genomic studies of human cancers have yielded a wealth of information about genes that are altered in tumors1,2,3. A challenge arising from these studies is that many genes are altered, and it can be difficult to distinguish genetic alterations that drove tumorigenesis from that those arose incidentally during transformation. To draw this distinction it is beneficial to have an assay that can quantitatively measure the effect of an altered gene on tumor initiation and other processes that enable tumors to persist and disseminate. Here we present a rapid means to screen large numbers of candidate melanoma modifiers in zebrafish using an autochthonous tumor model4 that encompasses steps required for melanoma initiation and maintenance. A key reagent in this assay is the miniCoopR vector, which couples a wild-type copy of the mitfa melanocyte specification factor to a Gateway recombination cassette into which candidate melanoma genes can be recombined5. The miniCoopR vector has a mitfa rescuing minigene which contains the promoter, open reading frame and 3'-untranslated region of the wild-type mitfa gene. It allows us to make constructs using full-length open reading frames of candidate melanoma modifiers. These individual clones can then be injected into single cell Tg(mitfa:BRAFV600E);p53(lf);mitfa(lf)zebrafish embryos. The miniCoopR vector gets integrated by Tol2-mediated transgenesis6 and rescues melanocytes. Because they are physically coupled to the mitfa rescuing minigene, candidate genes are expressed in rescued melanocytes, some of which will transform and develop into tumors. The effect of a candidate gene on melanoma initiation and melanoma cell properties can be measured using melanoma-free survival curves, invasion assays, antibody staining and transplantation assays.

A rapid way to screen for melanoma modifiers using a zebrafish autochthonous tumor model is presented. It takes advantage of the miniCoopR vector which allows for expression of candidate melanoma genes in melanocytes. A method to obtain melanoma-free survival curves, an invasion assay, a protocol for antibody staining of scale melanocytes and a melanoma transplantation assay are described.

Zebrafish Autochthonous 종양 모델을 사용하여 흑종 조절을위한 검사

Genomic studies of  human cancers have yielded a wealth of information about genes that are altered  in tumors1,2,3. A challenge arising from these ...

Construction of Vapor Chambers Used to Expose Mice to Alcohol During the Equivalent of all Three Trimesters of Human Development

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal Alcohol Spectrum Disorders (FASDs). At the most severe end of the spectrum is Fetal Alcohol Syndrome (FAS), characterized by growth retardation, craniofacial dysmorphology, and neurobehavioral deficits. Studies with animal models, including rodents, have elucidated many molecular and cellular mechanisms involved in the pathophysiology of FASDs. Ethanol administration to pregnant rodents has been used to model human exposure during the first and second trimesters of pregnancy. Third trimester ethanol consumption in humans has been modeled using neonatal rodents. However, few rodent studies have characterized the effect of ethanol exposure during the equivalent to all three trimesters of human pregnancy, a pattern of exposure that is common in pregnant women. Here, we show how to build vapor chambers from readily obtainable materials that can each accommodate up to six standard mouse cages. We describe a vapor chamber paradigm that can be used to model exposure to ethanol, with minimal handling, during all three trimesters. Our studies demonstrate that pregnant dams developed significant metabolic tolerance to ethanol. However, neonatal mice did not develop metabolic tolerance and the number of fetuses, fetus weight, placenta weight, number of pups/litter, number of dead pups/litter, and pup weight were not significantly affected by ethanol exposure. An important advantage of this paradigm is its applicability to studies with genetically-modified mice. Additionally, this paradigm minimizes handling of animals, a major confound in fetal alcohol research.

We demonstrate the construction of alcohol vapor chambers using readily available materials that simultaneously house 6 mouse cages. We further describe their use in a mouse model of fetal alcohol exposure equivalent to all 3&#160;trimesters of human pregnancy. This paradigm exposes animals during gestation and postnatal days 1-12.

인간 발달의 세 삼 분기의 상당 기간 동안 알코올에 쥐를 노출하는 데 사용 증기 챔버의 건설

Exposure to alcohol during development can result in a constellation of morphological and behavioral abnormalities that are collectively known as Fetal ...

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. Despite the discovery of key regulators of uterine contractility, this process is still not fully understood. Transgenic mice provide an ideal model in which to study parturition. Previously, the only method to study uterine contractility in the mouse was ex vivo isometric tension recordings, which are suboptimal for several reasons. The uterus must be removed from its physiological environment, a limited time course of investigation is possible, and the mice must be sacrificed. The recent development of radiometric telemetry has allowed for longitudinal, real-time measurements of in vivo intrauterine pressure in mice. Here, the implantation of an intrauterine telemeter to measure pressure changes in the mouse uterus from mid-pregnancy until delivery is described. By comparing differences in pressures between wild type and transgenic mice, the physiological impact of a gene of interest can be elucidated. This technique should expedite the development of therapeutics used to treat myometrial disorders during pregnancy, including preterm labor.

Intrauterine Telemetry to Measure Mouse Contractile Pressure In Vivo

This manuscript provides a protocol for implanting telemeters in the mouse for the purpose of measuring intrauterine pressures during pregnancy.

자궁 원격 측정은 마우스 수축 압력을 측정하는<em&gt; 생체</em

A complex integration of molecular and electrical signals is needed to transform a quiescent uterus into a contractile organ at the end of pregnancy. ...

Studying Pancreatic Cancer Stem Cell Characteristics for Developing New Treatment Strategies

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor initiation, metastasis and resistance to radio- and chemotherapy. Here we describe a specific methodology for culturing primary human pancreatic CSCs as tumor spheres in anchorage-independent conditions. Cells are grown in serum-free, non-adherent conditions in order to enrich for CSCs while their more differentiated progenies do not survive and proliferate during the initial phase following seeding of single cells. This assay can be used to estimate the percentage of CSCs present in a population of tumor cells. Both size (which can range from 35 to 250 micrometers) and number of tumor spheres formed represents CSC activity harbored in either bulk populations of cultured cancer cells or freshly harvested and digested tumors 1,2. Using this assay, we recently found that metformin selectively ablates pancreatic CSCs; a finding that was subsequently further corroborated by demonstrating diminished expression of pluripotency-associated genes/surface markers and reduced in vivo tumorigenicity of metformin-treated cells. As the final step for preclinical development we treated mice bearing established tumors with metformin and found significantly prolonged survival. Clinical studies testing the use of metformin in patients with PDAC are currently underway (e.g., NCT01210911, NCT01167738, and NCT01488552). Mechanistically, we found that metformin induces a fatal energy crisis in CSCs by enhancing reactive oxygen species (ROS) production and reducing mitochondrial transmembrane potential. In contrast, non-CSCs were not eliminated by metformin treatment, but rather underwent reversible cell cycle arrest. Therefore, our study serves as a successful example for the potential of in vitro sphere formation as a screening tool to identify compounds that potentially target CSCs, but this technique will require further in vitro and in vivo validation to eliminate false discoveries.

Pancreatic cancer stem cells (CSCs) can be expanded in vitro using the anchorage-independent sphere culture technique, which represents a powerful tool to study CSC biology and can serve as the first step to develop novel CSC-targeting therapies. Here the methodology for expanding, analyzing and targeting of pancreatic CSCs is provided.

췌장암 새로운 치료 전략을 개발하기위한 전지 특성 줄기 공부

Pancreatic ductal adenocarcinoma (PDAC) contains a subset of exclusively tumorigenic cancer stem cells (CSCs) which have been shown to drive tumor ...

Boldness, Aggression, and Shoaling Assays for Zebrafish Behavioral Syndromes

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is important to standardize protocols to ensure proper analyses and interpretations. In our previous studies, we have measured boldness by monitoring a series of behaviors (time near surface, latency in transitions, number of transitions, and darts) in a 1.5 L trapezoidal tank. Likewise, we quantified aggression by observing bites, lateral displays, darts, and time near an inclined mirror in a rectangular 19 L tank. By dividing a 76 L tank into thirds, we also examined shoaling preferences. The shoaling assay is a highly customizable assay and can be tailored for specific hypotheses. However, protocols for this assay also must be standardized, yet flexible enough for customization. In previous studies, end chambers were either empty, contained 5 or 10 zebrafish, or 5 pearl danios (D. albolineatus). In the following manuscript, we present a detailed protocol and representative data that accompany successful applications of the protocol, which will allow for replication of behavioral syndrome experiments.

This manuscript describes the setup, implementation, and analysis of boldness, aggression, and shoaling in zebrafish and testing for the presence of a behavioral syndrome. A standardized approach for behavioral quantification will allow for easier comparison across studies. Modifications to this protocol are possible as each assay can be run individually.

Zebrafish의 행동 증후군에 대한 대담, 침략, 그리고이 얕아 분석

A behavioral syndrome exists when specific behaviors interact under different contexts. Zebrafish have been test subjects in recent studies and it is ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

급성 폐 손상의 열기 기관 절개 위산 흡인 쥐 모델은 최대한 급성 치명적 폐 손상의 결과

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...

Conducting Maximal and Submaximal Endurance Exercise Testing to Measure Physiological and Biological Responses to Acute Exercise in Humans

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic responses to acute exercise are predominantly influenced by the duration and intensity of the exercise regimen. As exercise is increasingly thought of as a therapeutic treatment and/or diagnostic tool, it is important that standardizable methodologies be utilized to understand the variability and to increase the reproducibility of exercise outputs and measurements of responses to such regimens. To that end, we describe two different cycling exercise regimens that yield different physiologic outputs. In a maximal exercise test, exercise intensity is continually increased with a greater workload resulting in an increasing cardiopulmonary and metabolic response (heart rate, stroke volume, ventilation, oxygen consumption and carbon dioxide production). In contrast, during endurance exercise tests, the demand is increased from that at rest, but is raised to a fixed submaximal exercise intensity resulting in a cardiopulmonary and metabolic response that typically plateaus. Along with the protocols, we provide suggestions on measuring physiologic outputs that include, but are not limited to, heart rate, slow and forced vital capacity, gas exchange metrics, and blood pressure to enable the comparison of exercise outputs between studies. Biospecimens can then be sampled to assess cellular, protein, and/or gene expression responses. Overall, this approach can be easily adapted into both short- and long-term effects of two distinct exercise regimens.

To assess the influence of exercise intensity on physiologic and biologic responses, two different exercise testing protocols were utilized. Methods outlining exercise testing on a cycle ergometer as an incremental maximal oxygen consumption test and endurance, steady state submaximal endurance test are described.

최대한 하 고 Submaximal 지구력 운동 인간에서 급성 운동 생리 및 생물학 응답을 측정 하기 위해 테스트를 실시

Regular physical activity has a positive effect on human health, but the mechanisms controlling these effects remain unclear. The physiologic and biologic ...

Precision Implementation of Minimal Erythema Dose (MED) Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary to produce erythema (inflammatory reddening) on the surface of the skin. In this context, the MED is regarded as a key factor in determining starting doses for UV phototherapy for common skin conditions such as psoriasis and eczema. In research settings, MED testing also has potential to be a powerful tool for assessing within- and between-persons variation in inflammatory responses. However, MED testing has not been widely adopted for use in research settings, likely owing to a lack of published guidelines, which is a barrier to obtaining reproducible results from this assay. Also, protocols and equipment for establishing MED vary widely, making it difficult to compare results across laboratories. Here, we describe a precise and reproducible method to induce and measure superficial erythema using newly designed protocols and methods that can easily be adapted to other equipment and laboratory environments. The method described here includes detail on procedures that will allow extrapolation of a standardized dosage schedule to other equipment so that this protocol can be adapted to any UV radiation source.

Precision Implementation of Minimal Erythema Dose MED Testing to Assess Individual Variation in Human Inflammatory Response

Minimal erythema dose (MED) testing is used to establish dosage schedules for ultraviolet radiation phototherapy. It can assess individual variation in inflammatory response but lacks methodology for achieving reproducible results. Here, we present a precision implementation of MED and demonstrate its ability to capture individual variation in inflammatory response.

최소한의 홍반 복용량의 정밀 한 구현 (MED) 인간의 염증 반응에 개별 변화를 평가 하는 테스트

Minimal erythema dose (MED) testing is frequently used in clinical settings for determining the smallest amount of ultraviolet (UV) irradiation necessary ...

Murine Precision-Cut Liver Slices as an Ex Vivo Model of Liver Biology

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.

This protocol provides a simple and reliable method for the production of viable precision-cut liver slices from mice. The ex vivo tissue samples can be maintained under laboratory tissue culture conditions for multiple days, providing a flexible model to examine liver pathobiology.

뮤린 정밀 절단 간 슬라이스 간 생물학의 Ex Vivo 모델로

Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic ...