저자 아무 승인 있다.

동물 연구로 원리의 실험실 동물 관리 (건강의 국가 학회 간행물 출간: 8, 2011 년) 독일 입법에 의해 경 세 하는 규칙을 따라 했다. 공식 허가 정부 동물 관리 사무실 (Landesamt 위한 Natur, Umwelt und Verbraucherschutz 노르 트 베스트 팔 렌, Recklinghausen, 독일)에서 부여 됩니다.
1입니다. 동물
<ol><li>열린 감 금 소에 여성 독일 landrace 돼지 (25-30 kg 무게)를 사용 합니다.</li>
	<li>그룹 (아르곤과 헬륨) 당 5 동물을 사용 합니다.</li>
	<li>실험 하기 전에 적어도 1 주일에 대 한 환경에 순응을 동물 허용. 물에 대 한 무료 액세스와 함께 수술 이전 24 시간에 대 한 빠른 동물.</li>
</ol>2입니다. 마 취
<ol><li>Premedicate 동물 마 취 제 (15 mg/kg 몸 무게 [BW]), xylazine (10 mg/kg BW), 및 아트로핀 (0.1 mg/kg BW) 근육 주사와 마 취의 유도 하기 전에 10 분.</li>
	<li>주변 정 맥 접근 배치 22 게이지 정의 귀 정 맥에 의해 설정 됩니다.</li>
	<li>Propofol 2 mg/kg 체중의 i.v. 주입 하 여 전신 마 취를 유도.</li>
	<li>부정사 위치에 동물을 놓고 2 cm. 피하 조직의 무딘 준비를 통해 정 맥 찾기의 길이 jugular 홈에 경도 피부 절 개를 수행 합니다. 정, 다음 Seldinger 와이어를 삽입 합니다.</li>
	<li>정을 철회 하 고 가이드를 통해 14 백. 카 테 터를 삽입 합니다. 가이드 와이어를 철회. 카 테 터는 내선 번호로 연결 하 고 스트랩 또는 봉합 하 여 카 테 터를 흥분 시키는.</li>
	<li>바로 후 두 경 삽입를 혀 밖으로 당기고. 풀 다운은 후두개는 후 두 경의 팁을 사용 합니다. 성 대를 통해 튜브를 삽입 합니다. Glottis 아래 갑을 놓고 팽창 합니다.</li>
	<li>36와 42 mm Hg 사이 끝 갯벌 부분 탄소 이산화 긴장을 유지 하 20-26 호흡/min과 10 ml/kg의 해 일 볼륨에서 40% 산소로 환기.</li>
	<li>1-1.5%의 농도에서 isoflurane와 3-4 µ g/k g/h의 농도에 펜타닐 마 취를 유지 합니다.</li>
	<li>4 mL/kg의 초기 속도로 벨의 젖 산 솔루션 공급/h, 8 mL/kg의 일정 한 주입 속도에 개복 술 후 증가/h.</li>
</ol>3. 수술 및 플라즈마 응고
<ol><li>표준 수술 테이블에 부정사 위치에 동물을 배치 합니다.</li>
	<li>표준 외과 살 균 제를 적용 하 여 피부를 소독 (2-프로 판 올 45 g/100 g, 1-프로 판 올 10 g/100 g, 비페닐-2-ol 0.2 g/100 g) 3 번 수술 면봉으로.</li>
	<li>메스와 액을 넓은 중간 선 개복 술 칼 프로세스에서 수행 하 고 외과 견인 기를 설치.</li>
	<li>플라즈마 응고 장치에 스위치, 아르곤 또는 헬륨 가스 병 사용된 캐리어 가스에 따라 오픈. 원하는 대로 3 L/분 선택 응고 장치 출력 전력에 가스 흐름을 조정 합니다. 참고: 두 노블 가스, 아르곤 또는 헬륨, 플라즈마 응고에 대 한 사용할 수 있습니다. 응고 효과 비교할 수 있습니다. 자세한 내용은 참조7 를 참조 하십시오.</li>
	<li>앞에서 설명한7왼쪽된 간 엽에 플라즈마 응고를 수행 합니다. 티타늄 형 (사각 조리개 1 x 1 c m2)를 사용 하 여 응고 영역을 표준화. 5 응고 프로브 거리 1 cm의 s. 5 mm (그림 1)의 coagulations 사이의 짧은 거리와 다른 전원 설정을 coagulations는 수행할 수 있습니다.</li>
	<li>간의 수확에 대 한 모든 인 대 연결 간을 나눕니다. 분리 하 고 문맥 및 일반적인 담 관의 긴 부분을 떠나 우수한 십이지 장 굴곡 위 간 작은 꽃 자루를 나눕니다. 위와 간 아래 caval 정 맥을 분할 하 고 기관 검색.</li>
	<li>간, 수확 후 돼지 0.16 g/kg BW pentobarbital i.v. 관리에 의해 안락사 되었다.</li>
	<li>버스트 압력 실험에 대 한 날카로운가 위 왼쪽된 중간 간 엽의 절반을 resect. 플라즈마 응고 컷 (100W 출력) 표면 또는 섬유 소 실 란 트 (그림 2)와 함께 잘라 표면 밀봉.</li>
</ol>4. 미세 측정
참고: 레이저 도플러 분광학은 적혈구의 움직임으로 인 한 도플러 이동 측정을 통해 조직에 혈액 흐름을 확인할 수 있습니다. 레이저 신호 이동 적혈구의 수와 상관 한다. 레이저 도플러 분광학 임상 사용 (예: 이식 의학)과 여러 번15확인.
<ol><li>레이저 도플러 유량 계 및 분 광 광도 계에 전환 합니다. 플랫 프로브를 사용 합니다.</li>
	<li>흐름 및 속도 대 한 기준 측정을 가져가 라. 저장 하거나 값을 확인 합니다.</li>
	<li>3.5에서 설명 된 대로 응고를 수행 합니다.</li>
	<li>응고 사이트 및 측정 흐름 및 속도에 플랫 프로브를 배치 합니다. 다시 저장 하거나 값을 참고 합니다.</li>
	<li>Coagulator 장치의 모든 전원 설정에 대 한 반복 합니다.</li>
</ol>5. 온도 측정
<ol><li>(열감지 카메라, 노트북, 및 적외선 온도계)에 시스템을 전환 하 고 측정을 수행 하기 전에 적어도 1 시간 동안 실행 하자.</li>
	<li>초점을 조정 하 고 응고 사이트에 열감지 카메라에 프레임을 봅니다. 20 보다 더 큰 온도 해상도 1024 x 768 픽셀의 해상도와 적외선 시퀀스를 검출 될 수 있다 mK. 고려, 그 응고와 주변 조직의 지역-열 전달에 의해 영향을-보기 중간에 위치 하 고 있습니다. 참고: 그것은 가능한 최적 공간 해상도 대 한 프레임의 픽셀을 포함 해야 합니다.</li>
	<li>2 분 동안 열감지 카메라와 함께 간 표면에 플라즈마 coagulator과 응고 과정을 기록 합니다.</li>
	<li>온도 기록 분석 소프트웨어와 함께 이미지 시퀀스 분석: 관심의 영역을 정의. 참고: 소프트웨어 시간 해당 평균 온도 과정을 계산합니다.</li>
</ol>6. 응고 깊이 측정
<ol><li>날카로운가 위 왼쪽된 중간 간 엽 수확.</li>
	<li>1 cm 두께가 응고 사이트를 삭제할. 추가 처리를 위해 3 m m 두꺼운 경도 세그먼트를 잘라.</li>
	<li>밤새 중립 10%와 4 ° C에서 수정 조직 샘플 버퍼링 말린. 열 용융 점 이상 2 ° C 파라핀 하 고 분할 영역을 포함. 프로세스 하룻밤입니다.</li>
	<li>되며/오신 얼룩을 수행 합니다.<ol>
<li>Deparaffinize 및 이후 2에 담거 서 조직을 수 화 x 크 실 렌, 100% 에탄올 (EtOH), 95 %EtOH, 70 %EtOH, 이온된 H2O 2 분.</li>
		<li>3 분 위한 마이어의 되며 솔루션 조직 샘플을 얼룩.</li>
		<li>이제 5 분 동안 수돗물에 씻어.</li>
		<li>3 분 오신 솔루션 조직 얼룩.</li>
		<li>3 분 2 x EtOH 95% 그리고 크 실 렌에 린스. 표준 설치 매체와 함께 탑재.</li>
	</ol>
</li>
	<li>(현미경 연결 된 카메라, 이미징 소프트웨어)에 시스템을 전환 합니다. 40 X 확대와 함께 모든 섹션을 보기.</li>
	<li>40 X의 확대는 개체 마이크로미터의 이미지를 가져가 라. 재조정 목표 창에서 버튼을 누릅니다. 수동 보정을 선택 합니다. 100 µ m. 대화 상자에서 m m 입력 0, 1 마이크로미터 이미지에 라인을 그릴 하 고 확인을 누릅니다.</li>
	<li>보기에서 길이 선택 > 분석 컨트롤 > 주석 및 측정 창. 마우스로 응고 여백에 간 표면에서 측정 합니다. 내보내기 또는 결과 참고. 같은 슬라이드에 다른 위치에 측정을 반복 합니다. 참고: 응고 깊이 쉽게 정상적인 hepatocyte 코드와 수축 된 세포질, pyknotic 핵 및 출혈 영역 괴의 영역 사이의 날카로운 차이로 정상적인 간 조직에서 분화 될 수 있다.</li>
	<li>두 측정의 평균을 계산 합니다.</li>
</ol>7. 버스트 압력 측정
<ol><li>(자동 펌프, 압력 미터)에 시스템을 전환 합니다. 3.7 단계에 따라 간 견본을 준비 합니다. 참고: 3-방법-자 지를 통해 연결 된 두 개의 병렬 펌프를 사용 합니다. 1500 mm Hg의 최대 압력은 단일 펌프와 함께 얻을 수 없습니다.</li>
	<li>햅 틱 작은 꽃 자루가 위 문맥, 일반적인 간 동맥 및 담 즙 덕트를 격리 합니다. 문맥 overholt 집게와 클램프와 monofil 봉합 4-0와 선. 일반적인 간 동맥 overholt 집게를 죄 십시오 그리고 monofil 봉합 4-0와 선.</li>
	<li>일반적인 담 관으로 채널 16 카 테 터를 삽입 하 고 2-0 비단 봉합 사로 선. 자동 펌프 카 테 터를 연결, 압력 미터 (그림 3)와 함께 3 자 지를 설치 합니다.</li>
	<li>식 염 수로 관류 주사기를 채우십시오.</li>
	<li>99 mL/h의 배달 속도와 자동 펌프를 시작 합니다.</li>
	<li>버스트 압력 누설 및 기록에 대 한 잘라 간 표면과 압력 미터를 모니터링 합니다. 참고: 누설의 더 쉬운 인식을 위해 특허 블루는 식 염 수 (2 mL 특허 블루 + 18 mL 식 염 수)을 추가할 수 있습니다. 버스트 압력 압력 미터에 압력의 손실의 시간을 주의 하 여 관찰 하는 것이 쉽습니다.</li>
</ol>

<ol>
	<li>Link, W. J., Incropera, F. P., Glover, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+plasma+scalpel:+comparison+of+tissue+damage+and+wound+healing+with+electrosurgical+and+steel+scalpels.">A plasma scalpel: comparison of tissue damage and wound healing with electrosurgical and steel scalpels.</a> ArchSurg. 111, 392-397 (1976).</li><li>Kwon, A. H., Inui, H., Kamiyama, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Successful+laparoscopic+haemostasis+using+an+argon+beam+coagulator+for+blunt+traumatic+splenic+injury.">Successful laparoscopic haemostasis using an argon beam coagulator for blunt traumatic splenic injury.</a> EurJSurg. 167, 316-318 (2001).</li><li>Frilling, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effectiveness+of+a+new+carrier-bound+fibrin+sealant+versus+argon+beamer+as+haemostatic+agent+during+liver+resection:+a+randomised+prospective+trial.">Effectiveness of a new carrier-bound fibrin sealant versus argon beamer as haemostatic agent during liver resection: a randomised prospective trial.</a> Langenbecks ArchSurg. 390, 114-120 (2005).</li><li>Raiser, J., Zenker, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+plasma+coagulation+for+open+surgical+and+endoscopic+applications:+state+of+the+art.">Argon plasma coagulation for open surgical and endoscopic applications: state of the art.</a> J Phys Appl Phys. 39 (16), 3520-3523 (2006).</li><li>Farin, G., Grund, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Technology+of+argon+plasma+coagulation+with+particular+regard+to+endoscopic+applications.">Technology of argon plasma coagulation with particular regard to endoscopic applications.</a> EndoscSurgAllied Technol. 2, 71-77 (1994).</li><li>Grund, K. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+plasma+coagulation+(APC):+ballyhoo+or+breakthrough?.">Argon plasma coagulation (APC): ballyhoo or breakthrough?.</a> Endoscopy. 29, 196-198 (1997).</li><li>Glowka, T. R., Standop, J., Paschenda, P., Czaplik, M., Kalff, J. C., Tolba, R. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+and+helium+plasma+coagulation+of+porcine+liver+tissue.">Argon and helium plasma coagulation of porcine liver tissue.</a> J Int Med Res. , (2017).</li><li>Dowling, R. D., Ochoa, J., Yousem, S. A., Peitzman, A., Udekwu, A. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Argon+beam+coagulation+is+superior+to+conventional+techniques+in+repair+of+experimental+splenic+injury.">Argon beam coagulation is superior to conventional techniques in repair of experimental splenic injury.</a> JTrauma. 31, 717-720 (1991).</li><li>Go, P. M., Goodman, G. R., Bruhn, E. W., Hunter, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+argon+beam+coagulator+provides+rapid+hemostasis+of+experimental+hepatic+and+splenic+hemorrhage+in+anticoagulated+dogs.">The argon beam coagulator provides rapid hemostasis of experimental hepatic and splenic hemorrhage in anticoagulated dogs.</a> JTrauma. 31, 1294-1300 (1991).</li><li>Brand, C. U., Blum, A., Schlegel, A., Farin, G., Garbe, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+argon+plasma+coagulation+in+skin+surgery.">Application of argon plasma coagulation in skin surgery.</a> Dermatology. 197, 152-157 (1998).</li><li>Carus, T., Rackebrandt, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Collateral+tissue+damage+by+several+types+of+coagulation+(monopolar,+bipolar,+cold+plasma+and+ultrasonic)+in+a+minimally+invasive,+perfused+liver+model.">Collateral tissue damage by several types of coagulation (monopolar, bipolar, cold plasma and ultrasonic) in a minimally invasive, perfused liver model.</a> ISRNSurg. , 518924 (2011).</li><li>Bludau, M., Vallbohmer, D., Gutschow, C., Holscher, A. H., Schroder, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+measurement+of+gastric+mucosal+microcirculation+using+a+combined+laser+Doppler+flowmeter+and+spectrophotometer.">Quantitative measurement of gastric mucosal microcirculation using a combined laser Doppler flowmeter and spectrophotometer.</a> DisEsophagus. , (2008).</li><li>Beckert, S., Witte, M. B., Konigsrainer, A., Coerper, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+impact+of+the+Micro-Lightguide+O2C+for+the+quantification+of+tissue+ischemia+in+diabetic+foot+ulcers.">The impact of the Micro-Lightguide O2C for the quantification of tissue ischemia in diabetic foot ulcers.</a> Diabetes Care. 27, 2863-2867 (2004).</li><li>Erdogan, D., de Graaf, W., van Gulik, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesive+strength+of+fibrinogen-coated+collagen+patch+or+liquid+fibrin+sealant+in+an+experimental+liver+resection+model+in+pigs.">Adhesive strength of fibrinogen-coated collagen patch or liquid fibrin sealant in an experimental liver resection model in pigs.</a> Eur Surg Res Eur Chir Forsch Rech Chir Eur. 41 (3), 298-302 (2008).</li><li>Knobloch, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microcirculation+of+the+sternum+following+harvesting+of+the+left+internal+mammary+artery.">Microcirculation of the sternum following harvesting of the left internal mammary artery.</a> ThoracCardiovascSurg. 51, 255-259 (2003).</li><li>Kanzler, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recommendation+for+severity+assessment+following+liver+resection+and+liver+transplantation+in+rats:+Part+I.">Recommendation for severity assessment following liver resection and liver transplantation in rats: Part I.</a> Lab Anim. 50 (6), 459-467 (2016).</li><li>Pehb&#246;ck, D., Dietrich, H., Klima, G., Paal, P., Lindner, K. H., Wenzel, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia+in+swine+optimizing+a+laboratory+model+to+optimize+translational+research.">Anesthesia in swine optimizing a laboratory model to optimize translational research.</a> Anaesthesist. 64 (1), 65-70 (2015).</li><li>Nykonenko, A., V&#225;vra, P., Zon&#269;a, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anatomic+Peculiarities+of+Pig+and+Human+Liver.">Anatomic Peculiarities of Pig and Human Liver.</a> Exp Clin Transplant Off J Middle East Soc Organ Transplant. 15 (1), 21-26 (2017).</li><li>Fechner, G., von Pezold, J., Luzar, O., Hauser, S., Tolba, R. H., M&#252;ller, 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=Modified+spectrometry+(O2C+device)+of+intraoperative+microperfusion+predicts+organ+function+after+kidney+transplantation:+a+pilot+study.">Modified spectrometry (O2C device) of intraoperative microperfusion predicts organ function after kidney transplantation: a pilot study.</a> Transplant Proc. 41 (9), 3575-3579 (2009).</li><li>Patterson, E. J., Scudamore, C. H., Owen, D. A., Nagy, A. G., Buczkowski, A. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiofrequency+ablation+of+porcine+liver+in+vivo:+effects+of+blood+flow+and+treatment+time+on+lesion+size.">Radiofrequency ablation of porcine liver in vivo: effects of blood flow and treatment time on lesion size.</a> AnnSurg. 227, 559-565 (1998).</li></ol>

저자는 공개 없다.

설치류 모델 간 수술에 대 한 오랜 시간16에 대 한 설정 됩니다. 그럼에도 불구 하 고, 큰 동물 모델 특정 이점을 제공:로 적용할 수 있는 인 간에 대 한 표준 수술 장비, 수술 기법 비교 임상 사용 표준 임상 평가 방법 수 없음 microsurgical 장비 필요 전송 실험. 예를 들어 표준 임상 혈액 검사 특별 한 실험실 테스트 방법 (그림 10)에 대 한 필요 없이 수행할 수 있...

아르곤 플라즈마 응고 (APC) 3 년간1,2보다 더 복 부 수술에서 널리 사용 되는 악기 이다. 그것은 이차 hemostasis의 성취에 대 한 표준 기술 간 씰링 하 여 주요 hepatectomy 잘라 나중 출혈3을 방지 하기 위해 표면. 플라즈마 응고는 이온화 가스의 아크를 통해 전기 에너지를 전달 하는 고주파 electrocautery의 특수 형태 이다. Monopolar electrothermal hemostasis, 제공이 비접촉 기술 조직4에 충실 하는 전극 방지의 이점이 있다. 이온화 가스 빔 자동으로 낮은 전기 저항 지역으로 이동 하 고 아직 포장해 다른 지역에 건조로 인해 저항 상승 때 멀리 설정 되어. 이 응고5,6의 균일 한 제한 된 깊이 생성합니다. 응고 효과 영향을 미치는 요소는 활성화 시간, 응고 장치 및 조직에는 프로브에서 거리의 전원 설정. 헬륨은 플라즈마 응고7사용할 수 있는 다른 캐리어 가스 이다. 최근 임상 연구에 집중된 임상 결과 보다는 조직학 및 기능적인 결과3,8,9, 체 외에서 조사10에 초점을 맞춘 실험 연구 하면서 또는 격리 끼얹는다 장기11에 실험.
기본 프로토콜 돼지에 인간 표준 장비를 사용 하 여 임상 응용에 가까운 큰 동물 모델에서 플라즈마 응고의 효과의 연구를 수 있습니다: 레이저 도플러 유량 계에 의해 미세 noninvasively 평가 및 분 광 광도 계,이 표시12,13표준 임상 도구입니다. 응고 하는 동안 온도 변화를 적외선 온도계와 열감지 카메라 모니터링 된다. 조직학 되며/오신의 조직 샘플의 수확 후 섹션 스테인드 응고의 깊이 측정 됩니다. 이차 hemostasis에 대 한 다른 방법 가진 비교에 대 한 파열 압력 실험 수행 됩니다. 앞에서 설명한 기법14, 달리 이러한 혈압 제외 explanted 장기에 실시 하는 효과 관련. 플라즈마 응고의 로컬 효과에 설명된 조사, 뿐만 아니라 돼지 모델에서 또한 표준 혈액 검사를 맡아 수 있다.

<table border="0" cellpadding="0" cellspacing="0" width="824">
	<tbody>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Xylazine 20 mg/mL</td>
			<td style="width:184px;">Vetoquinol GmbH</td>
			<td style="width:192px;">Xylapan</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Ketamine 100 mg/mL</td>
			<td style="width:184px;">Ceva GmbH</td>
			<td style="width:192px;">Ceva Ketamine Injection</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Atropine 100 mg / 10 mL</td>
			<td style="width:184px;">Dr. Franz K&#246;hler Chemie GmbH</td>
			<td style="width:192px;">Atropinsulfat K&#246;hler 100mg Amp.</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Propofol</td>
			<td style="width:184px;">Fresenius Kabi GmbH</td>
			<td style="width:192px;">Propofol 1% MCT Fresenius</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Fentanyl</td>
			<td style="width:184px;">KG Rotexmedica GmbH</td>
			<td style="width:192px;">Fentanyl 0,5mg Rotexmedica</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Isoflurane</td>
			<td style="width:184px;">Abbot GmbH</td>
			<td style="width:192px;">Forene 100% (V/V) 250 mL</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Ringer&#39;s lactate solution</td>
			<td style="width:184px;">Baxter Deutschland GmbH</td>
			<td style="width:192px;"></td>
			<td style="width:232px;">sodium 131mmol/l, potassium 5 mmol/l, calcium 2 mmol/l, cloride 111 mmol/l, lactate 29 mmol/l</td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Surgical disinfactant</td>
			<td style="width:184px;">Sch&#252;lke &#38; Mayr GmbH</td>
			<td style="width:192px;">Kodan Tinktur forte gef&#228;rbt 1l 104804</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Motorized microscope</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">Eclipse TE2000-E</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Microscope camera</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">Digitalsight DS-Qi1Mc</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Imaging software</td>
			<td style="width:184px;">Nikon Instruments Europe</td>
			<td style="width:192px;">NIS elements Vers. 4.40</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Plasma coagulator</td>
			<td style="width:184px;">S&#246;ring GmbH</td>
			<td style="width:192px;">CPC-1000</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Argon gas</td>
			<td style="width:184px;">Linde AG</td>
			<td style="width:192px;">Argon 4.8&#160;</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Helium gas</td>
			<td style="width:184px;">Linde AG</td>
			<td style="width:192px;">Helium 4.8</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">O2C</td>
			<td style="width:184px;">LEA Medizintechnik GmbH</td>
			<td style="width:192px;">O2C Version 1212</td>
			<td style="width:232px;">with LF-2 or LF-3 probe</td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;">Infrared thermometer</td>
			<td style="width:184px;">Voltcraft</td>
			<td>VOLTCRAFT IR 260-8S</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Thermographic camera</td>
			<td>InfraTec GmbH</td>
			<td>VarioCAM HD head 820</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Thermographic analysis sofrtware</td>
			<td>InfraTec GmbH</td>
			<td style="width:192px;">IRBIS 3</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Mayer&#39;s Hematoxylin solution</td>
			<td></td>
			<td style="width:192px;">Merck 1.09249</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Eosin solution</td>
			<td style="width:184px;">VWR International GmbH</td>
			<td style="width:192px;">Merck 1.09844</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Rollerpump Masterflex L/S easy Load</td>
			<td style="width:184px;">Cole-Parmer Instrument Company</td>
			<td style="width:192px;">model 7518-10</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusorpump</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">Perfusor secura FT</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Digital pressure meter</td>
			<td style="width:184px;">Greisinger electronic</td>
			<td style="width:192px;">GMH 3161</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusorsyringe, 50 mL</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 8728810 F</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Perfusor line, Type IV Standard, PVC Luer lock</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 8722960</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">3-Way stopcock, Dicofix C35C</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;">REF 16494 C</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Silk 2-0. 3 metric</td>
			<td style="width:184px;">Resorba</td>
			<td style="width:192px;">REF H5F</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">Vicryl 4-0 Sutupak</td>
			<td style="width:184px;">Ethicon</td>
			<td>V1224H</td>
			<td style="width:232px;"></td>
		</tr>
		<tr height="47">
			<td height="47" style="height:47px;width:216px;">NaCl 0.9 %</td>
			<td style="width:184px;">B. Braun Melsungen AG</td>
			<td style="width:192px;"></td>
			<td style="width:232px;"></td>
		</tr>
	</tbody>
</table>

assessment of plasma coagulation on liver tissue in a large animal model in vivo

플라즈마 응고 electrocautery의 형태로 나중 단계에서 출혈을 방지 하기 위해 주요 hepatectomy 후 큰 간 잘라 표면 밀봉 수십 년 간 수술에 사용 됩니다. 간 조직에 플라즈마 응고의 정확한 효과 가난 하 게 검사 합니다. 우리의 돼지 모델에서 응고 효과 임상 응용 프로그램에 가까운 시험 될 수 있다. 결합 된 레이저 도플러 유량 계 및 분 광 광도 계 문서 미세 변경 8 m m 조직 깊이에서 응고 하는 동안 noninvasively, 주관적인 임상 인상 넘어 hemostasis에 대 한 정량 정보를 제공 합니다. 응고 사이트에서 온도 응고 하는 동안 적외선 온도계 사전 및 사후 응고와 열감지 카메라 평가, 가스 빔 온도 측정 소자의 상한값 때문에 가능 하지 않습니다. 응고의 깊이 되며/오신 교정 후 개체 마이크로미터 섹션을 물 들일에 현미경으로 측정 되 고 전원 설정-응고 깊이 관계에 대 한 정확한 정보를 제공. 그것은 큰 혈관을 밀봉 하기 위하여 플라즈마 coagulator 불가능으로 씰링 효과 담 관에 시험 된다. 버스트 압력 실험 수행 됩니다 밖으로 배제 혈압 explanted 장기 효과 관련.

미세: 플라즈마 응고 다음 hemostasis 위한 진단 장치를 활용 하 여 미세 혈관 변화에 의해 설명할 수 있습니다. 142.7 ± 76.08의 기준 값에서 모 세관 혈액 흐름 (임의의 단위 (AU)로 표시) 감소 57.78 ± 49.57 하는 누구나 48.5 ± 7.26에 25 W 장치 출력 전력, AU AU 50 W와 5.04 ± 1.31 AU 100 W (그림 4).
온도:</...

Watch this Scientific Journal Video about Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo at JoVE.com

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

여기 선물이 비보에간 조직에 플라즈마 응고를 실험적으로 평가 하는 프로토콜. 돼지 모델에서 미세 레이저 도플러 검사, 응고 깊이 조직학 측정, 온도 적외선 온도계와 열감지 카메라와 덕트 씰링 효과 의해 응고 사이트에서 버스트 압력에 의해 문서화 실험입니다.

플라즈마 응고는 큰 동물 모델에서 에서 Vivo에서 간 조직에의 평가

Plasma coagulation as a form of electrocautery is used in liver surgery for decades to seal the large liver cut surface after major hepatectomy to prevent ...

assessment-plasma-coagulation-on-liver-tissue-large-animal-model

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Medicine

Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

8.8K 조회수.  University of Bonn. 여기 선물이 비보에간 조직에 플라즈마 응고를 실험적으로 평가 하는 프로토콜. 돼지 모델에서 미세 레이저 도플러 검사, 응고 깊이 조직학 측정, 온도 적외선 온도계와 열감지 카메라와 덕트 씰링 효과 의해 응고 사이트에서 버스트 압력에 의해 문서화 실험입니다.

비디오: Assessment of Plasma Coagulation on Liver Tissue in a Large Animal Model In Vivo

An Isolated Working Heart System for Large Animal Models

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart model, have been invaluable tools for studying cardiovascular function and disease1-15. Although the Langendorff heart preparation can be used for any mammalian heart, most studies involving this apparatus use small animal models (e.g., mouse, rat, and rabbit) due to the increased complexity of systems for larger mammals1,3,11. One major difficulty is ensuring a constant coronary perfusion pressure over a range of different heart sizes &#8211; a key component of any experiment utilizing this device1,11. By replacing the classic hydrostatic afterload column with a centrifugal pump, the Langendorff working heart apparatus described below allows for easy adjustment and tight regulation of perfusion pressures, meaning the same set-up can be used for various species or heart sizes. Furthermore, this configuration can also seamlessly switch between constant pressure or constant flow during reperfusion, depending on the user&#8217;s preferences. The open nature of this setup, despite making temperature regulation more difficult than other designs, allows for easy collection of effluent and ventricular pressure-volume data.

Most studies involving the Langendorff apparatus use small animal models due to the increased complexity of systems for larger mammals. We describe a Langendorff system for large animal models that allows for use across a range of species, including humans, and relatively easy data acquisition.

큰 동물 모델에 대한 격리 된 작업 심장 시스템

Since its introduction in the late 19th century, the Langendorff isolated heart perfusion apparatus, and the subsequent development of the working heart ...

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Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic patient tumors in different assays1. Human tumor xenografts represent the gold standard with respect to tumor biology, drug discovery, and metastasis research 2-4. Tumor xenografts can be derived from different types of material like tumor cell lines, tumor tissue from primary patient tumors4 or serially transplanted tumors. When propagated in vivo, xenografted tissue is infiltrated and vascularized by cells of mouse origin. Multiple factors such as the tumor entity, the origin of xenografted material, growth rate and region of transplantation influence the composition and the amount of mouse cells present in tumor xenografts. However, even when these factors are kept constant, the degree of mouse cell contamination is highly variable.
Contaminating mouse cells significantly impair downstream analyses of human tumor xenografts. As mouse fibroblasts show high plating efficacies and proliferation rates, they tend to overgrow cultures of human tumor cells, especially slowly proliferating subpopulations. Mouse cell derived DNA, mRNA, and protein components can bias downstream gene expression analysis, next-generation sequencing, as well as proteome analysis 5. To overcome these limitations, we have developed a fast and easy method to isolate untouched human tumor cells from xenografted tumor tissue. This procedure is based on the comprehensive depletion of cells of mouse origin by combining automated tissue dissociation with the benchtop tissue dissociator and magnetic cell sorting. Here, we demonstrate that human target cells can be can be obtained with purities higher than 96% within less than 20 min independent of the tumor type.

Human tumor xenografts are vascularized and infiltrated by cells of mouse origin during the growth phase in vivo. To circumvent the bias caused by these contaminating cells during downstream analysis, we developed a method allowing for comprehensive depletion of all mouse cells by magnetic cell sorting.

인간 종양 이종 이식에서 마우스 세포의 고갈은 크게 대상 세포의 다운 스트림 분석을 향상

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

Assessment of Child Anthropometry in a Large Epidemiologic Study

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to understand health effects of child growth and adiposity. Gold standard methods of measuring adiposity, such as dual X-ray absorptiometry (DXA), may not be feasible in large field studies. Research staff can, however, complete anthropometric measurements, such as body circumferences and skinfold measurements, using inexpensive portable equipment. In this protocol we detail how to obtain manual anthropometric measurements from children, including standing and sitting height, weight, waist circumference, hip circumference, mid-upper arm circumference, triceps skinfold thickness, and subscapular skinfold thickness, and procedures to assess the quality of these measurements. To demonstrate accuracy of these measurements, among 1,110 school-aged children in the pre-birth cohort Project Viva we calculated Spearman correlation coefficients comparing manual anthropometric measurements with a gold standard measure of body fat, DXA fat mass1. To address reliability, we evaluate intra-rater technical error of measurement at a quality control session conducted on adult female volunteers.

In epidemiologic studies of children, well-trained research staff can accurately and precisely assess weight, height, sitting height, skinfold thicknesses, and body circumferences.

A high proportion of children have overweight and obesity in the United States and other countries. Accurate assessment of anthropometry is essential to ...

Surgical Angiogenesis in Porcine Tibial Allotransplantation: A New Large Animal Bone Vascularized Composite Allotransplantation Model

Segmental bone loss resulting from trauma, infection malignancy and congenital anomaly remains a major reconstructive challenge. Current therapeutic options have significant risk of failure and substantial morbidity.
Use of bone vascularized composite allotransplantation (VCA) would offer both a close match of resected bone size and shape and the healing and remodeling potential of living bone. At present, life-long drug immunosuppression (IS) is required. Organ toxicity, opportunistic infection and neoplasm risks are of concern to treat such non-lethal indications.
We have previously demonstrated that bone and joint VCA viability may be maintained in rats and rabbits without the need of long-term-immunosuppression by implantation of recipient derived vessels within the VCA. It generates an autogenous, neoangiogenic circulation with measurable flow and active bone remodeling, requiring only 2 weeks of IS. As small animals differ from man substantially in anatomy, bone physiology and immunology, we have developed a porcine bone VCA model to evaluate this technique before clinical application is undertaken. Miniature swine are currently widely used for allotransplantation research, given their immunologic, anatomic, physiologic and size similarities to man. Here, we describe a new porcine orthotopic tibial bone VCA model to test the role of autogenous surgical angiogenesis to maintain VCA viability.
The model reconstructs segmental tibial bone defects using size- and shape-matched allogeneic tibial bone segments, transplanted across a major swine leukocyte antigen (SLA) mismatch in Yucatan miniature swine. Nutrient vessel repair and implantation of recipient derived autogenous vessels into the medullary canal of allogeneic tibial bone segments is performed in combination with simultaneous short-term IS. This permits a neoangiogenic autogenous circulation to develop from the implanted tissue, maintaining flow through the allogeneic nutrient vessels for a short time. Once established, the new autogenous circulation maintains bone viability following cessation of drug therapy and subsequent nutrient vessel thrombosis.

Currently any kind of vascularized composite allotransplantation depends on long-term-immunosuppression, difficult to support for non-life-critical indications. We present a new porcine tibial VCA model that can be used to study bone VCA and demonstrate the use of surgical angiogenesis to maintain bone viability without the need of long-term immune-modulation.

돼지 Tibial Allotransplantation 외과 신생: 새로운 큰 동물 뼈 나가도록 복합 Allotransplantation 모델

Segmental bone loss resulting from trauma, infection malignancy and congenital anomaly remains a major reconstructive challenge. Current therapeutic ...

Invasive Hemodynamic Monitoring of Aortic and Pulmonary Artery Hemodynamics in a Large Animal Model of ARDS

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to pulmonary hypertension. For a better understanding and treatment of this disease, precise hemodynamic monitoring of left and right ventricular parameters is important. For this reason, it is essential to establish experimental pig models of cardiac hemodynamics and measurements for research purpose. 
This article shows the induction of ARDS by using oleic acid (OA) and consequent right ventricular dysfunction, as well as the instrumentation of the pigs and the data acquisition process that is needed to assess hemodynamic parameters. To achieve right ventricular dysfunction, we used oleic acid (OA) to cause ARDS and accompanied this with pulmonary artery hypertension (PAH). With this model of PAH and consecutive right ventricular dysfunction, many hemodynamic parameters can be measured, and right ventricular volume load can be detected. 
All vital parameters, including respiratory rate (RR), heart rate (HR) and body temperature were recorded throughout the whole experiment. Hemodynamic parameters including femoral artery pressure (FAP), aortic pressure (AP), right ventricular pressure (peak systolic, end systolic and end diastolic right ventricular pressure), central venous pressure (CVP), pulmonary artery pressure (PAP) and left arterial pressure (LAP) were measured as well as perfusion parameters including ascending aortic flow (AAF) and pulmonary artery flow (PAF). Hemodynamic measurements were performed using transcardiopulmonary thermodilution to provide cardiac output (CO). Furthermore, the PiCCO2 system (Pulse Contour Cardiac Output System 2) was used to receive parameters such as stroke volume variance (SVV), pulse pressure variance (PPV), as well as extravascular lung water (EVLW) and global end-diastolic volume (GEDV). Our monitoring procedure is suitable for detecting right ventricular dysfunction and monitoring hemodynamic findings before and after volume administration.

We present a protocol of creating right ventricular dysfunction in a pig model by inducing ARDS. We demonstrate invasive monitoring of left and right ventricular cardiac output using flow probes around the aorta and the pulmonary artery, as well as blood pressure measurements in the aorta and pulmonary artery.

대동맥 및 폐 동맥 Hemodynamics 아즈의 큰 동물 모델에서의 침략 Hemodynamic 감시

One of the leading causes of morbidity and mortality in patients with heart failure is right ventricular (RV) dysfunction, especially if it is due to ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, normothermic ex vivo liver perfusion (NEVLP) has been introduced as a method to evaluate and modify organ function. NEVLP has many advantages in comparison to hypothermic and subnormothermic perfusion including reduced preservation injury, restoration of normal organ function under physiologic conditions, assessment of organ performance, and as a platform for organ repair, remodeling, and modification. Both murine and porcine NEVLP models have been described. We demonstrate a rat model of NEVLP and use this model to show one of its important applications &#8212; the use of a therapeutic molecule added to liver perfusate. Catalase is an endogenous reactive oxygen species (ROS) scavenger and has been demonstrated to decrease ischemia-reperfusion in the eye, brain, and lung. Pegylation has been shown to target catalase to the endothelium. Here, we added pegylated-catalase (PEG-CAT) to the base perfusate and demonstrated its ability to mitigate liver preservation injury. An advantage of our rodent NEVLP model is that it is inexpensive in comparison to larger animal models. A limitation of this study is that it does not currently include post-perfusion liver transplantation. Therefore, prediction of the function of the organ post-transplantation cannot be made with certainty. However, the rat liver transplant model is well established and certainly could be used in conjunction with this model. In conclusion, we have demonstrated an inexpensive, simple, easily replicable NEVLP model using rats. Applications of this model can include testing novel perfusates and perfusate additives, testing software designed for organ evaluation, and experiments designed to repair organs.

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

There is a significant liver donor shortage, and criteria for liver donors have been expanded. Normothermic ex vivo liver perfusion (NEVLP) has been developed to evaluate and modify organ function. This study demonstrates a rat model of NEVLP and tests the ability of pegylated-catalase, to mitigate liver preservation injury.

Ex Vivo Normothermic 간 관류의 작은 동물 모델

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, we have translated a vascular injury model from a small animal into a large animal (canine), with slight modifications that can be used for pre-clinical screening of prophylactic and thrombolytic agents. In addition to the surgical methods, the modified protocol describes the step-by-step methods to assess carotid artery canalization by angiography, detailed instructions to process both the brain and carotid artery for histological analysis to verify carotid canalization and cerebral hemorrhage, and specific parameters to complete an assessment of downstream thromboembolic events by utilizing magnetic resonance imaging (MRI). In addition, specific procedural changes from the previously well-established small animal model necessary to translate into a large animal (canine) vascular injury are discussed.

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

제 2 철 염화 유도 개 경 동맥 혈전 증: 혈관 손상의 큰 동물 모델

Occlusive arterial thrombosis leading to cerebral ischemic stroke and myocardial infarction contributes to ~13 million deaths every year globally. Here, ...

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 ...

In Vitro Model of Coronary Angiogenesis

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects in these vessels represent severe health risks such as in atherosclerosis, which can lead to myocardial infarctions and heart failures in patients. Consequently, coronary artery disease is one of the leading causes of death worldwide. Despite its clinical importance, relatively little progress has been made on how to regenerate damaged coronary arteries. Nevertheless, recent progress has been made in understanding the cellular origin and differentiation pathways of coronary vessel development. The advent of tools and technologies that allow researchers to fluorescently label progenitor cells, follow their fate, and visualize progenies in vivo have been instrumental in understanding coronary vessel development. In vivo studies are valuable, but have limitations in terms of speed, accessibility, and flexibility in experimental design. Alternatively, accurate in vitro models of coronary angiogenesis can circumvent these limitations and allow researchers to interrogate important biological questions with speed and flexibility. The lack of appropriate in vitro model systems may have hindered the progress in understanding the cellular and molecular mechanisms of coronary vessel growth. Here, we describe an in vitro culture system to grow coronary vessels from the sinus venosus (SV) and endocardium (Endo), the two progenitor tissues from which many of the coronary vessels arise. We also confirmed that the cultures accurately recapitulate some of the known in vivo mechanisms. For instance, we show that the angiogenic sprouts in culture from SV downregulate COUP-TFII expression similar to what is observed in vivo. In addition, we show that VEGF-A, a well-known angiogenic factor in vivo, robustly stimulates angiogenesis from both the SV and Endo cultures. Collectively, we have devised an accurate in vitro culture model to study coronary angiogenesis.

In vitro models of coronary angiogenesis can be utilized for the discovery of the cellular and molecular mechanisms of coronary angiogenesis. In vitro explant cultures of sinus venosus and endocardium tissues show robust growth in response to VEGF-A and display a similar pattern of COUP-TFII expression as in vivo.

관상 동맥 혈관 신생의 체외 모델

Here, we describe an in vitro culture assay to study coronary angiogenesis. Coronary vessels feed the heart muscle and are of clinical importance. Defects ...

Using a Chemical Biopsy for Graft Quality Assessment

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated with an increased survival rate and greater quality of patient&#8217;s life when compared to conventional dialysis. Regrettably, transplantology suffers from a lack of reliable methods for organ quality assessment. Standard diagnostic techniques are limited to macroscopic appearance inspection or invasive tissue biopsy, which do not provide comprehensive information about the graft. The proposed protocol aims to introduce solid phase microextraction (SPME) as an ideal analytical method for comprehensive metabolomics and lipidomic analysis of all low molecular compounds present in kidneys allocated for transplantation. The small size of the SPME probe enables performance of a chemical biopsy, which enables extraction of metabolites directly from the organ without any tissue collection. The minimum invasiveness of the method permits execution of multiple analyses over time: directly after organ harvesting, during its preservation, and immediately after revascularization at the recipient&#8217;s body. It is hypothesized that the combination of this novel sampling method with a high-resolution mass spectrometer will allow for discrimination of a set of characteristic compounds that could serve as biological markers of graft quality and indicators of possible development of organ dysfunction.

The protocol presents utilization of the chemical biopsy approach followed by comprehensive metabolomic and lipidomic analysis for quality assessment of kidney grafts allocated for transplantation.

이식 품질 평가를 위해 화학 생검 사용

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...