Name: pseudofracture: an acute peripheral tissue trauma model
Uploaded: 2011-04-18T11:00:00
Description: Watch this Scientific Journal Video about Pseudofracture: An Acute Peripheral Tissue Trauma Model at JoVE.com

출혈성 쇼크 GM053789의 기금 소스 / 번호 분자 생물학

1. 악기, 외과 필드 준비 : 모든 실험 절차는 무균 기술을 사용하여 수행됩니다. 시작하기 전에 실험 지역은 철저하게 청소하고 소독해야합니다. benchtop은 공기 건조 후 70 % 알코올로 닦여 수, 소독해야합니다. 실험 작업 영역에 드레싱 수술 블루 패드 및 살균 필드를 배치합니다. 모든 자료와 악기 압력솥 사용하기 전에 소독을하고 있습니다. 주사기와 바?...

<ol>
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F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+alterations+in+plasma+IL-6+and+TNF+levels+after+trauma+and+hemorrhage.">Differential alterations in plasma IL-6 and TNF levels after trauma and hemorrhage.</a> Am J Physiol. 260, R167-R171 (1991).</li><li>Kalicke, T., Schlegel, U., Printzen, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+a+standardized+closed+soft+tissue+trauma+on+resistance+to+local+infection.+An+experimental+study+in+rats.">Influence of a standardized closed soft tissue trauma on resistance to local infection. An experimental study in rats.</a> J Ortho Res. 21, 373-378 (2003).</li><li>Kobbe, P., Vodovotz, Y., Kaczorowski, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+fracture+associated+soft+tissue+injury+in+the+induction+of+sytemic+inflammation+and+remote+organ+dysfunction+after+bilateral+femur+fracture.">The role of fracture associated soft tissue injury in the induction of sytemic inflammation and remote organ dysfunction after bilateral femur fracture.</a> J Ortho Trauma. 22, 385-390 (2008).</li><li>Kobbe, P., Vodovotz, Y., Kaczorwoski, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pattern+of+cytokine+release+and+evolution+of+remote+organ+dysfunction+after+bilateral+femur+fracture.">Pattern of cytokine release and evolution of remote organ dysfunction after bilateral femur fracture.</a> Shock. 30, 43-47 (2008).</li><li>Flohe, S. B., Flohe, S., Schade, F. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deterioration+of+the+immune+system+after+trauma:+signals+and+cellular+mechanisms.">Deterioration of the immune system after trauma: signals and cellular mechanisms.</a> Inn. Immun. 14, 333-344 (2008).</li><li>Maier, B., LeFering, R., Lhenert, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+versus+late+onset+of+multiple+organ+failure+is+associated+with+differing+patterns+of+plasma+cytokine+biomarker+expression+and+outcome+after+severe+trauma.">Early versus late onset of multiple organ failure is associated with differing patterns of plasma cytokine biomarker expression and outcome after severe trauma.</a> Shock. 28, 668-674 (2007).</li><li>Mestas, J., Hughes, C. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Of+mice+not+men:+differences+between+mouse+and+human+immunology.">Of mice not men: differences between mouse and human immunology.</a> J. Immunol. 172, 2731-2738 (2004).</li><li>Hoth, J. J., Wells, J. D., Brownlee, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toll+like+receptor+4-dependent+responses+to+lung+injury+in+a+murine+model+of+pulmonary+contusion.">Toll like receptor 4-dependent responses to lung injury in a murine model of pulmonary contusion.</a> Shock. 31, 376-381 (2009).</li><li>Matsutani, T., Samy, A. n. a. n. t. h. a., Rue, T. S., W, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transgenic+prolactin-/-+mice:+effect+of+trauma-hemorrhage+on+splenocyte+functions.">Transgenic prolactin-/- mice: effect of trauma-hemorrhage on splenocyte functions.</a> Am J Physiol Cell Physiol. 288, 1109-1116 (2005).</li><li>Matsutani, T., Samy, A. n. a. n. t. h. a., Kang, T. S., S-C, . <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+genetic+background+influences+severity+of+immune+responses+following+trauma-hemorrhage.">Mouse genetic background influences severity of immune responses following trauma-hemorrhage.</a> Cytokine. 30, 168-117 (2005).</li><li>Tsukamoto, T., Pape, H. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemorrhagic+shock+in+endotoxin+resistant+mice+:+Improved+survival+unrelated+to+deficient+production+of+tumor+necrosis+factor.">Hemorrhagic shock in endotoxin resistant mice : Improved survival unrelated to deficient production of tumor necrosis factor.</a> J Trauma. 35, 720-724 (1993).</li><li>Jamsa, T., Jalovaara, P., Peng, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+three-point+bending+test+and+peripheral+quantitative+computed+tomography+analysis+in+the+evaluation+of+the+strength+of+mouse+femur.">Comparison of three-point bending test and peripheral quantitative computed tomography analysis in the evaluation of the strength of mouse femur.</a> Bone. 23, 155-161 (1998).</li><li>Bounarens, F., Einhorn, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+a+standard+closed+fracture+in+laboratory+animal+bone.">Production of a standard closed fracture in laboratory animal bone.</a> J Ortho Res. 2, 97-101 (1984).</li><li>Holstein, J. H., Menger, M. D., Culemann, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+a+locking+femur+nail+for+mice.">Development of a locking femur nail for mice.</a> J Biomech. 40, 215-219 (2007).</li><li>Holstein, J. 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P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+a+closed+femur+fracture+model+in+mice.">Characterization of a closed femur fracture model in mice.</a> J Ortho Trauma. 18, 687-695 (2004).</li><li>Mark, H., Bergholm, J., Nilsson, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+external+fixation+method+and+device+to+study+fracture+healing+in+rats.">An external fixation method and device to study fracture healing in rats.</a> Acta Orthop Scand. 74, 476-487 (2003).</li><li>Bhandari, M., Shanghnessy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+minimally+invasive+percutaneous+technique+of+intramedullary+nail+insertion+in+an+animal+model+of+fracture+healing.">A minimally invasive percutaneous technique of intramedullary nail insertion in an animal model of fracture healing.</a> Arch Orthop Trauma Surg. 121, 591-593 (2001).</li><li>Sonanis, S. V., Lampard, A. L., Kamat, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simple+technique+to+remove+a+bent+femoral+intramedullary+nail+and+broken+interlocking+screw.">A simple technique to remove a bent femoral intramedullary nail and broken interlocking screw.</a> J Trauma. 63, 435-438 (2007).</li><li>Kobbe, P., Kaczorwoski, D. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluid+resuscitation:+past,+present,+and+the+future.">Fluid resuscitation: past, present, and the future.</a> Shock. 33, 229-241 (2010).</li><li>Hierholzer, C., Billiar, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+in+the+early+phase+of+hemorrhagic+shock.">Molecular mechanisms in the early phase of hemorrhagic shock.</a> Lang Arch Surg. 386, 302-308 (2001).</li><li>Chaudry, I., Ayala, A., Ertel, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hemorrhage+and+resuscitation:+immunological+aspects.">Hemorrhage and resuscitation: immunological aspects.</a> Am J Physiol. 259, 63-678 (1990).</li><li>Kawasaki, T., Hubbard, W. J., Choudhry, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Trauma-hemorrhage+induces+depressed+splenic+dendritic+cell+functions+in+mice.">Trauma-hemorrhage induces depressed splenic dendritic cell functions in mice.</a> J Immunol. 177, 4514-4520 (2006).</li><li>Kohut, L., Darwiche, S. S., Frank, A. M., Brumfield, J. M., Billiar, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fixed+volume+or+Fixed+Pressure:+A+Murine+Model+of+Hemorrhgaic+Shock.">Fixed volume or Fixed Pressure: A Murine Model of Hemorrhgaic Shock.</a> J Vis Exp. , (2010).</li><li>Menzel, C. L., Pfeifer, R., Darwiche, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+of+lower+extremity+damage+in+mice:+time+course+of+organ+damage+and+immune+response.">Models of lower extremity damage in mice: time course of organ damage and immune response.</a> J Surg Res. 166, e149-e156 (2011).</li><li>Pfeifer, R., Kobbe, P., Darwiche, S. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+hemorrhage+in+the+induction+of+systemic+inflammation+and+remote+organ+damage:+Analysis+of+combined+pseudo-fracture+and+hemorrhagic+shock.">Role of hemorrhage in the induction of systemic inflammation and remote organ damage: Analysis of combined pseudo-fracture and hemorrhagic shock.</a> J Orthop Res. 29, 270-274 (2011).</li></ol>

Pseudofracture, 무균 musculoskeletal 외상의 재현성 murine 모델, 외상후 면역 반응의 평가를 위해 수 있습니다. pseudofracture 모델 immunologically 긴 골절의 기능의 레크 리에이션을 통해 끝단 파괴 환경을 모방 : 부상 근육과 부드러운 조직은 원래 뼈 한방울 흘리지 않고 손상된 뼈 및 골수에 노출되는 38,39 biphasic 면역 반응은 다음과 같은 볼 수 있습니다. 48hrs 주변 여물로 묘사 지연 immunosuppression의 두 ?...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Surgical blue pad</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>50-7105</td>
<td>&nbsp;</td>
<tr>
<td>Sterile Field dressings</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>NC9517505</td>
<td>&nbsp;</td>
<tr>
<td>Circulating heating pad 18"x26"</td>
<td>&nbsp;</td>
<td>Harvard</td>
<td>py872-5272</td>
<td>&nbsp;</td>
<tr>
<td>Hot bead instrument sterilizer</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>11156-002</td>
<td>&nbsp;</td>
<tr>
<td>Stainless steel tray 8" x 11"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>62687-049</td>
<td>&nbsp;</td>
<tr>
<td>Plexiglass boards (10x15x0.5cm) </td>
<td>&nbsp;</td>
<td>University of Pittsburgh Machine shop</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Tape rolls 1"</td>
<td>&nbsp;</td>
<td>Corporate Express</td>
<td>MMM26001</td>
<td>&nbsp;</td>
<tr>
<td>50cc conical tube</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Straight side wide mouth jars (used as cap for nose cone)</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>159000-058</td>
<td>&nbsp;</td>
<tr>
<td>Oster A5 clippers w. size 40 blade</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>10749-020</td>
<td>&nbsp;</td>
<tr>
<td>Surgical scissors (straight &#x2013; 12cm)</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>14068-12</td>
<td>&nbsp;</td>
<tr>
<td>Hemostats curved -18cm</td>
<td>&nbsp;</td>
<td>Harvard</td>
<td>81331718</td>
<td>&nbsp;</td>
<tr>
<td>Forceps (0.8mm-tip, curved-10cm)</td>
<td>&nbsp;</td>
<td>Fine Science Tools</td>
<td>11050-10</td>
<td>&nbsp;</td>
<tr>
<td>Gauze 4"x4"</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1.5cc microfuge tube</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Ice bucket</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Mortar and Pestle </td>
<td>&nbsp;</td>
<td>Fisher</td>
<td>12-961AA</td>
<td>&nbsp;</td>
<tr>
<td>1cc syringe w/ 25G needle</td>
<td>&nbsp;</td>
<td>Fisher Scientific</td>
<td>14-826-88</td>
<td>&nbsp;</td>
<tr>
<td>20G needle</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1mL pipetteman</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>1mL pipette tips</td>
<td>&nbsp;</td>
<td>can be purchased through any global vendor</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Falcon polystyrene 8ml tubes </td>
<td>&nbsp;</td>
<td>VWR</td>
<td>60819-331</td>
<td>&nbsp;</td>
<tr>
<td>Sterilization pouch 3"x8"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>24008</td>
<td>&nbsp;</td>
<tr>
<td>Sterilization pouch 5"x10"</td>
<td>&nbsp;</td>
<td>VWR</td>
<td>24010</td>
<td>&nbsp;</td>
<tr>
<td>MacConkey II Agar plate</td>
<td>&nbsp;</td>
<td>BD Biosciences</td>
<td>221172</td>
<td>&nbsp;</td>
<tr>
<td>Ethyl Alcohol - 200 proof </td>
<td>&nbsp;</td>
<td>Pharmaco-AAPER</td>
<td>&nbsp;</td>
<td>[70%]</td>
</tr>
<tr>
<td>Pentobarbital Sodium (Nembutol Sodium Solution)</td>
<td>&nbsp;</td>
<td>Ovation </td>
<td>&nbsp;</td>
<td>70mg/kg</td>
</tr>
<tr>
<td>Aerrane (Isoflurane)</td>
<td>&nbsp;</td>
<td>Baxter </td>
<td>&nbsp;</td>
<td>99.9%</td>
</tr>
<tr>
<td>Triadine Povidone Iodine (Betadine)</td>
<td>&nbsp;</td>
<td>Triad disposables</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Phosphate Buffered Saline (PBS)</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Buprenorphine HCl</td>
<td>&nbsp;</td>
<td>Bedford Laboratories</td>
<td>&nbsp;</td>
<td>0.1mg/kg</td>
</tr>		</tbody>
		</table>
		</div>

pseudofracture: an acute peripheral tissue trauma model

외상 다음 여러 장기의 장애와 외상 환자에서 높은 사망률을 초래할 수있는 초기 하이퍼 반응 염증 반응이있다,이 반응은 종종 감염의 임상적 합병증을 추가 지연 immunosuppression 동반되며 사망률을 높일 수 있습니다 1-9가. 많은 연구 외상 다음 면역 체계의 반응에서 이러한 변화를 평가하기 시작했습니다. 10-15도 Immunologic 연구는 크게 유전자 변형 및 생체내 모델링에 사용할 수 녹아웃 마우스의 다양한 통해 지원되며. 자세한 조사에서 이러한 변종이 원조는 immunologic 응답에 관련된 분자 경로를 평가할 16-21 마우스의 파괴 고정 기술이 복잡하고 쉽게 재현할 수는 없습니다로서 실험 murine 외상 모델링에 도전, 장기 조사입니다. 22-30 마취의 지속, 장기간 사용하지 않고 동물과 장기 생존에 이동의 자유를 허용하면서이 pseudofracture 모델, 쉽게 복제 외상 모델은, immunologically 끝단 파괴 환경을 흉내낸하여 이러한 어려움을 극복. 목적은 긴 골절의 기능을 재현하는 것입니다, 부상 근육과 부드러운 조직은 원래 뼈 한방울 흘리지 않고 손상된 뼈 및 골수에 노출되어 있습니다. hindlimbs에 양자 근육 호감 부상이 부상을 근육으로 뼈를 솔루션의 주입에 의해 다음 : pseudofracture 모델은 두 부분으로 구성되어 있습니다. 본 솔루션은 연령과 체중 일치 syngeneic 기증자 모두 hindlimbs에서 긴 뼈가 수확하여 준비가되어 있습니다. 이 유골은 다음 조각과 인산염의 resuspended하는 것은 뼈 솔루션을 만들 수 살린 버퍼. 양자 대퇴골 골절은 끝단 외상의 일반적으로 사용하고 잘 구축 모델이며, pseudofracture 모델의 개발 기간 동안 비교 모델. 우리가 멸균 아직 비례 심한 말초 조직 외상 모델을 원하는 사용 가능한 파괴 모델의 다양한 가운데, 우리는 pseudofracture 우리의 비교와 같은 부드러운 조직 손상과 파괴의 폐쇄 방법을 사용하기로 결정했습니다. 31 출혈성 쇼크가 심각한 외상의 설정에서 일반적인 발견이며, 세계 hypoperfusion는 외상 모델에 매우 관련된 요소를 추가합니다. 32-36 pseudofracture 모델이 쉽게 높은 심각도의 여러 외상 모델 출혈성 쇼크 모델과 결합 수 있습니다 . 37

Watch this Scientific Journal Video about Pseudofracture: An Acute Peripheral Tissue Trauma Model at JoVE.com

Pseudofracture: An Acute Peripheral Tissue Trauma Model

Pseudofracture, 무균 musculoskeletal 외상의 재현성 murine 모델은, 늦은 용어 외상후 면역 반응의 평가를 위해 수 있습니다. 이 문서는 여러 외상의 연구를 허용하는 실험 모델 조합의 가능성을 포함하여 단계 모델 단계의 절차 실행을 설명합니다.

Pseudofracture : 급성 주변 조직 외상 모델

Following trauma there is an early hyper-reactive inflammatory response that can lead to multiple organ dysfunction and high mortality in trauma patients; ...

pseudofracture-an-acute-peripheral-tissue-trauma-model

Research

JoVE Journal

Medicine

Expired CO2 Measurement in Intubated or Spontaneously Breathing Patients from the Emergency Department

Carbon dioxide (CO2) along with oxygen (O2) share the role of being the most important gases in the human body. The measuring of expired CO2 at the mouth has solicited growing clinical interest among physicians in the emergency department for various indications: (1) surveillance et monitoring of the intubated patient; (2) verification of the correct positioning of an endotracheal tube; (3) monitoring of a patient in cardiac arrest; (4) achieving normocapnia in intubated head trauma patients; (5) monitoring ventilation during procedural sedation. The video allows physicians to familiarize themselves with the use of capnography and the text offers a review of the theory and principals involved. In particular, the importance of CO2 for the organism, the relevance of measuring expired CO2, the differences between arterial and expired CO2, the material used in capnography with their artifacts and traps, will be reviewed. Since the main reluctance in the use of expired CO2 measurement is due to lack of correct knowledge concerning the physiopathology of CO2 by the physician, we hope that this explanation and the video sequences accompanying will help resolve this limitation.

Expired CO2 Measurement in Intubated or Spontaneously Breathing Patients from the Emergency Department

This video describes how to perform CO2 measurement in intubated as well as spontaneously breathing patients. The main clinical indications refer to emergency situations: (1) verifying adequate positioning of an endotracheal tube; (2) achieving normocapnia in trauma patients; (3) monitoring ventilation in the case of procedural sedation.

만료 CO 2</sub응급실에서 Intubated 또는 자연스럽게 호흡 환자> 측정

Carbon dioxide (CO2) along with oxygen (O2) share the role of being the most important gases in the human body. The measuring of expired CO2 at the mouth ...

연구

의학

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of the hepatic blood supply is poorly oxygenated portal vein blood derived from the gastrointestinal tract and spleen. Oxygen is delivered to hepatocytes by blood flowing from a terminal branch of the portal vein to a central venule via sinusoids, and this makes an oxygen gradient in hepatic lobules. The oxygen gradient is an important physical parameter that involves the expression of enzymes upstream and downstream in hepatic microcirculation, but the lack of techniques for measuring oxygen consumption in the hepatic microcirculation has delayed the elucidation of mechanisms relating to oxygen metabolism in liver. We therefore used FITC-labeled erythrocytes to visualize the hepatic microcirculation and used laser-assisted phosphorimetry to measure the partial pressure of oxygen in the microvessels there. Noncontact and continuous optical measurement can quantify blood flow velocities, vessel diameters, and oxygen gradients related to oxygen consumption in the liver. In an acute hepatitis model we made by administering acetaminophen to mice we observed increased oxygen pressure in both portal and central venules but a decreased oxygen gradient in the sinusoids, indicating that hepatocyte necrosis in the pericentral zone could shift the oxygen pressure up and affect enzyme expression in the periportal zone. In conclusion, our optical methods for measuring hepatic hemodynamics and oxygen consumption can reveal mechanisms related to hepatic disease.

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

급성 간염 모델 신청 : 시각화와 간장 Microcirculation의 혈류와 산소 소비 분석

There is a considerable discrepancy between oxygen supply and demand in the liver because hepatic oxygen consumption is relatively high but about 70% of ...

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

경쟁 손쥐 골수 이식 모델에서 기증자 세포 Engraftment를 확인하려면 수량 실시간 PCR을 사용하여

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no effective treatment for it yet. Limitations of currently used animal models impede understanding of the pathology mechanism. Multiphoton microscopy allows studying cells and tissues within intact animal brains longitudinally under physiological and pathological conditions. Here, we describe two models of acute brain injury studied by means of two-photon imaging of brain cell behavior under posttraumatic conditions. A selected brain region is injured with a sharp needle to produce a trauma of a controlled width and depth in the brain parenchyma. Our method uses stereotaxic prick with a syringe needle, which can be combined with simultaneous drug application. We propose that this method can be used as an advanced tool to study cellular mechanisms of pathophysiological consequences of acute trauma in mammalian brain in vivo. In this video, we combine acute brain injury with two preparations: cranial window and skull thinning. We also discuss advantages and limitations of both preparations for multisession imaging of brain regeneration after trauma.

Acute brain trauma is a severe injury that has no adequate treatment to date. Multiphoton microscopy allows studying longitudinally the process of acute brain trauma development and probing therapeutical strategies in rodents. Two models of acute brain trauma studied with in vivo two-photon imaging of brain are demonstrated in this protocol.

세로 두 광자 이미징 다음 마우스의 급성 뇌 외상

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Ischemic Tissue Injury in the Dorsal Skinfold Chamber of the Mouse: A Skin Flap Model to Investigate Acute Persistent Ischemia

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are still occurring, particularly in reconstructive flap surgery. Multiple experimental flap models have been developed to analyze underlying causes and mechanisms and to investigate treatment strategies to prevent ischemic complications. The limiting factor of most models is the lacking possibility to directly and repetitively visualize microvascular architecture and hemodynamics. The goal of the protocol was to present a well-established mouse model affiliating these before mentioned lacking elements. Harder et al. have developed a model of a musculocutaneous flap with a random perfusion pattern that undergoes acute persistent ischemia and results in ~50% necrosis after 10 days if kept untreated. With the aid of intravital epi-fluorescence microscopy, this chamber model allows repetitive visualization of morphology and hemodynamics in different regions of interest over time. Associated processes such as apoptosis, inflammation, microvascular leakage and angiogenesis can be investigated and correlated to immunohistochemical and molecular protein assays. To date, the model has proven feasibility and reproducibility in several published experimental studies investigating the effect of pre-, peri- and postconditioning of ischemically challenged tissue.

The window of the murine dorsal skinfold chamber presented visualizes a zone of acute persistent ischemia of a musculocutaneous flap. Intravital epi-fluorescence microscopy permits for direct and repetitive assessment of the microvasculature and quantification of hemodynamics. Morphologic and hemodynamic results can further be correlated with histological and molecular analyses.

마우스의 등쪽 피하 지방 상공 회의소에서 허혈성 조직 손상 : 피부 플랩 모델은 급성 영구 허혈을 조사하기

Despite profound expertise and advanced surgical techniques, ischemia-induced complications ranging from wound breakdown to extensive tissue necrosis are ...

Direct Mouse Trauma/Burn Model of Heterotopic Ossification

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical procedures. The majority of animal models used to study HO rely on the application of exogenous substances, such as bone morphogenetic protein (BMP), exogenous cell constructs, or genetic mutations in BMP signaling. While these models are useful they do not accurately reproduce the inflammatory states that cause the majority of cases of HO. Here we describe a burn/tenotomy model in mice that reliably produces focused HO. This protocol involves creating a 30% total body surface area partial thickness contact burn on the dorsal skin as well as division of the Achilles tendon at its midpoint. Relying solely on traumatic injury to induce HO at a predictable location allows for time-course study of endochondral heterotopic bone formation from intrinsic physiologic processes and environment only. This method could prove instrumental in understanding the inflammatory and osteogenic pathways involved in trauma-induced HO. Furthermore, because HO develops in a predictable location and time-course in this model, it allows for research to improve early imaging strategies and treatment modalities to prevent HO formation.

An Achilles tenotomy and burn injury model of heterotopic ossification allows for the reliable study of trauma induced ectopic bone formation without the application of exogenous factors.

직접 마우스 외상 / 이소성 골화의 모델을 굽기

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical ...

An Acute Retinal Model for Evaluating Blood Retinal Barrier Breach and Potential Drugs for Treatment

A low-cost, easy-to-use and powerful model system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach. An inflammatory factor, histamine, is demonstrated to compromise vessel integrity in the cultured retina through positive staining of IgG outside of the blood vessels. The effects of histamine itself and those of candidate drugs for potential treatments, such as lipoxin A4, are assessed using three parameters: blood vessel leakage via IgG immunostaining, activation of M&#252;ller cells via GFAP staining and change in neuronal dendrites through staining for MAP2. Furthermore, the layered organization of the retina allows a detailed analysis of the processes of M&#252;ller and ganglion cells, such as changes in width and continuity. While the data presented is with swine retinal culture, the system is applicable to multiple species. Thus, the model provides a reliable tool to investigate the early effects of compromised retinal vessel integrity on different cell types and also to evaluate potential drug candidates for treatment.

A low-cost, easy-to-use and powerful system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach induced by histamine. Blood vessel leakage, M&#252;ller cell activation and the continuity of neuronal processes are utilized to assess the damage response and its reversal with a potential drug, lipoxin A4.

치료를 위해 혈액 망막 장벽 위반 및 잠재적 인 마약을 평가하는 급성 망막 모델

A low-cost, easy-to-use and powerful model system is established to evaluate potential treatments that could ameliorate blood retinal barrier breach. An ...

A Saline/Bipolar Radiofrequency Energy Device As an Adjunct for Hemostasis in Solid Organ Injury/Trauma

Solid organ (liver, spleen, and kidney) hemorrhage is often life-threatening and can be difficult to stop in critically ill patients. Traditional techniques for arresting this ongoing bleeding include coagulation by high voltage electrocautery, topical hemostatic application, and the delivery of ignited argon gas. The goal of this study/video was to demonstrate the efficacy of a new energy device for arresting persistent solid organ hemorrhage.A novel instrument utilizing bipolar radiofrequency (RF) energy which acts to ignite/boil dripping saline from a simple handpiece is employed to arrest ongoing bleeding from solid organ injuries in a porcine model. This instrument is extrapolated from experience within elective hepatic resections. An escalating series of injuries to solid organs within a porcine model will be created. This will be followed by arresting hemorrhage with this novel energy device in sequence. A standard suction device will also be employed. This simple saline/RF energy instrument has the potential to arrest ongoing solid organ surface/capsular bleeding, as well as moderate hemorrhage associated with deep lacerations.

The goal of this publication is to demonstrate the potential application of a novel device using simulated solid organ injuries in a porcine model.

솔리드 장기 손상/외상에서 Hemostasis에 대 한 부겸으로 식염수/양극성 무선 주파수 에너지 장치

Solid organ (liver, spleen, and kidney) hemorrhage is often life-threatening and can be difficult to stop in critically ill patients. Traditional ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and drugs. It is widely used in many studies on the physiological functions of vascular smooth muscle, the pathogenesis of vascular diseases such as hypertension, and the development of smooth muscle relaxant drugs. The mouse is a widely used model animal with a large pool of disease models and genetically modified strains. We introduced this method to measure the isometric contraction of mouse mesenteric artery in detail. A 1.4-mm segment of mouse mesenteric resistance artery was isolated and mounted on a myograph chamber by passing two steel wires through its lumen. After equilibration and normalization steps, the vessel segment was potentiated by a high-K+ solution twice prior to the contraction assay. As an example of the application of this method in drug development, we measured the relaxant effect of a novel natural substance, neoliensinine, isolated from a Chinese herb, embryos of the lotus seed (Nelumbo nucifera Gaertn.) on mouse mesenteric arteries. The vessel segments mounted on the myograph chamber were stimulated with a high-K+ solution. When the force tension reached a stable sustained phase, cumulative doses of neoliensinine were added to the chamber. We found that neoliensinine had a dose-dependent relaxant effect on smooth muscle contraction, thus suggesting that it bears potential activity against hypertension. In addition, as the vessel segment can survive at least 4 hours after mounting and maintain contractility induced by the high-K+ solution for many times, we suggest that the wire myograph system may be used for the time-consuming process of drug screening.

The wire myograph technique is used to investigate vascular smooth muscle functions and screen new drugs. We report a detailed protocol for measuring the isometric contractility of the mouse mesenteric artery and for screening new relaxants of vascular smooth muscle.

Myography 와이어를 사용 하 여 마우스 Mesenteric 동맥의 isometric 수축 측정

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

An Ex Vivo Tissue Culture Model for Fibrovascular Complications in Proliferative Diabetic Retinopathy

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No current animal models of diabetes and oxygen-induced retinopathy develop the full-range progressive changes manifested in human proliferative diabetic retinopathy (PDR). Therefore, understanding of the disease pathogenesis and pathophysiology has relied largely on the use of histological sections and vitreous samples in approaches that only provide steady-state information on the involved pathogenic factors. Increasing evidence indicates that dynamic cell-cell and cell-extracellular matrix (ECM) interactions in the context of three-dimensional (3D) microenvironments are essential for the mechanistic and functional studies towards the development of new treatment strategies. Therefore, we hypothesized that the pathological fibrovascular tissue surgically excised from eyes with PDR could be utilized to reliably unravel the cellular and molecular mechanisms of this devastating disease and to test the potential for novel clinical interventions. Towards this end, we developed a novel method for 3D ex vivo culture of surgically-excised patient-derived fibrovascular tissue (FT), which will serve as a relevant model of human PDR pathophysiology. The FTs are dissected into explants and embedded in fibrin matrix for ex vivo culture and 3D characterization. Whole-mount immunofluorescence of the native FTs and end-point cultures allows thorough investigation of tissue composition and multicellular processes, highlighting the importance of 3D tissue-level characterization for uncovering relevant features of PDR pathophysiology. This model will allow the simultaneous assessment of molecular mechanisms, cellular/tissue processes and treatment responses in the complex context of dynamic biochemical and physical interactions within the PDR tissue architecture and microenvironment. Since this model recapitulates PDR pathophysiology, it will also be amenable for testing or developing new treatments.

Here, we present a protocol to study the pathophysiology of proliferative diabetic retinopathy by using patient-derived, surgically-excised, fibrovascular tissues for three-dimensional native tissue characterization and ex vivo culture. This ex vivo culture model is also amenable for testing or developing new treatments.

증식 성 당뇨 망막 증에 Fibrovascular 합병증에 대 한 전 비보 조직 문화 모델

Diabetic retinopathy (DR) is the most common microvascular complication of diabetes and one of the leading causes of blindness in working-age adults. No ...