This work was supported by grant 13IRG14560023 from the American Heart Association and NIH grants AI078972 and HL108054 to CJO. Support for AOB was from the NIH National Center for Advancing Translational Sciences NIH ULTR001120, and F31 A110417701.

참고 : 모든 마우스 실험을 검토하고 샌 안토니오 텍사스 건강 과학 센터의 대학 (프로토콜 # 1 13032-34-01C)에서 기관 동물 케어 및 사용위원회에 의해 승인되었다. 동물 관리 및 공법 89-544 (동물 복지 법) 및 그 수정, 보건 서비스 지침 및 실험 동물의 관리 및 사용을위한 설명서 (보건 복지부의 미국학과)에 부착 된 실험 프로토콜을 지원합니다. 1. 감염 <ol><li> 10-12 주 나이에 세 사이의 남성과 여성 모두의 BALB / c 마우스를 가져옵니다. 참고 : S. 폐렴 구균 혈청 형 4 변형 TIGR4 16 염기 서열 된 악성 임상 격리입니다. </li><li> 광학 밀도 (OD) 0.5 (620)에 도달 할 때까지 5 % CO 2에서 37 ° C에서 토드 - 휴이 국물 TIGR4 성장 (1.0 × 108 콜로니 형성 단위 [CFU] / ㎖에 상응 함). (3)에서 10 분 동안 원심 분리에 의해 펠렛 배양 폐렴500 XG 및 진공 라인을 사용하여 상층 액을 제거합니다. 일시 및 1.0 × 104 CFU / ml의 최종 농도로 멸균 인산 완충 식염수 (PBS)로 희석 된 세균. </li><li> 유도 챔버를 사용하여, 산소 2.5 % 증발 이소 플루 란과 쥐를 마취. 무딘 핀셋과 부드러운 발가락 핀치 마취 상태를 확인합니다. <ol><li> 한 손으로는 목덜미에 의해 마취 마우스를 잡고 똑바로하면, S. 100 ㎕와 각 마우스 복강 (IP)을 주입 (1.0 × 103 CFU의 도전 량에 해당) 27 ~ 30 게이지 바늘과 주사기를 사용 폐렴 서스펜션. 케이지에서 마우스 다시 놓습니다. 마우스는 일반적으로 주사 후 30 ~ 40 초 각성. </li></ol></li><li> 감염이 적어도 (초) 24 또는 30 (고급) 시간 동안 할 수있게합니다. <ol><li> CO 2 질식하여 쥐를 안락사. 마우스를 보장하기 위해 자궁 경부 전위를 수행하는 사망했다. 안락사는 더 제거에 의해 확인단계 1.6의 마음. </li></ol></li><li> 알코올 면봉으로 꼬리를 소독하고 2-3 밀리미터 부분을 싹둑. 혈액의 2 μl를 수집하고 직렬 나트륨 헤파린 1 U / ml의 다섯 배를 포함하는 PBS에 혈액을 10 배 희석. 플레이트 직렬 트립신 간장 혈액 한천 플레이트에 희석과 식민지에서 다음날 추정 해 균혈증의 수준을 계산 CO 2 5 %에서 37 ° C에서 밤새 품어. </li><li> 수술 플랫폼에서 부정사 위치에 마우스를 고정. 70 % 에탄올과 팻 건조와 가슴 스프레이. 수술 가위와 집게가 흉강을 열고 사용, 갈비뼈를 제거하고 심장과 폐를 노출하는 격막을 가로로 쪼개다. 사용 가위은 심장에 연결된 혈관을 잘라 부드럽게 집게와 멍 않도록주의 마음을 절제. </li><li> PBS과 마음을 씻어 후 관상 섹션 조직 절편 동안 획득 될 수 있도록 조직 표본 수집 카세트에 배치합니다. 파라핀 삽입, 놀이터 용파라핀 임베딩 보내 10 % 완충 포르말린 용액과 다음 날에 카세트 CE. <ol><li> 또한 드라이 아이스에 배치 된 cryomold 내에서의 OCT 화합물을 이용하여 플래시 동결. </li></ol></li></ol> 심장 병변 및 병변 내 폐렴 구균 2. 시각화 <ol><li> 마음의 4 챔버는 각 조직 섹션에서 볼 수 있도록 파라핀 심장 부분을 잘라. 5 μm의 두께에 슬라이드를 잘라. </li><li> 표준 방법 (17)를 사용하여 헤 마톡 실린 및 에오신 (H &amp; E)와 Deparaffinize와 얼룩 슬라이드. </li><li> 배율 :보기 H &amp; E는 낮은 (100X, 200X) 및 높은 (400 배, 1000 배 기름 침지)에서 광학 현미경을 사용하여 심장 섹션 스테인드. 심근에 microlesions의 존재에 대해 마음을 특성화. 여기에, 100X 1.3 개구 계획-NEOFLUAR 목표를 장착 한 자이스 Axioskop이 현미경을 사용하여 H &amp; E 이미지를 수집. 참고 : 조기 심장 리터 스테이지esions는 면역 세포, 단핵구 것으로 보인다 무엇 (단핵구와 과립구)의 존재 자신의 차 색깔 모양으로 어떤 경우에 차별된다. 특히 30 시간에서 본 고급 병변에서 면역 세포가 발견하고 큰 공포 병변이 자신의 자리에서 관찰되어 있습니다. 이것은 심장 병변 개발 초기 단계 면역 세포의 유입이 발생하면서 요구 것으로 나타나지 않음에 유의하는 것이 중요하다. </li></ol> Microlesions 내 폐렴 구균 3. 면역 형광 현미경 <ol><li> 양으로 대전 된 유리 슬라이드 위에 5 μm의 장소 섹션의 두께로 마이크로톰와 cryomolds에 제 마음입니다. 얼어 붙은 심장 섹션 해동 자연 건조하도록 허용합니다. (25 ° C에서의 pH 6.8-7.2) 10 분 ~ 10 % 중성 포르말린에 대한 부분을 수정합니다. </li><li> 다음 PBS로 다시 씻어, PBS에서 0.2 % 트리톤 X-100에서 15 분 동안 Permeabilize 하시려면, 5 분 동안 PBS에서 슬라이드 (3 배)를 씻으십시오. </li><li> 차단 티슈 섹션에 ì<html> 1 시간 동안 PBS에서 10 % 염소 혈청과 슬라이드에. </li><li> PBS, 커버 슬라이드를 씻어 1 토끼 항 혈청 형 4 폐렴 구균 항혈청과 심장 부분을 품어 : 1,000 PBS에서 10 % 염소 혈청과 2 시간 동안 37 ° C에서. 참고 : 음성 대조군의 경우 연구자들은 순진한 토끼 항혈청을 사용한다 </li><li> 37 ° C에서 30 분 동안 2,000 : 1 염소 항 - 토끼 FITC 결합 된 항체와 10 % 염소 혈청 PBS로 섹션을 PBS, 커버로 세척하고 배양 한 후. </li><li> 제조업체의 지시에 따라, 진핵 세포의 핵의 시각화를 위해 5 ㎎ / ㎖에서 DAPI (4 &#39;, 6 Diamidino -2- 페닐 인돌)를 사용합니다. 씻고 FluorSave과 조직 섹션을 탑재합니다. </li><li> 낮고 높은 배율에서 모두 공 촛점 현미경 시스템을 사용하여 형광 이미지를 획득. 최적의 결과를 수득하는데 따라 개구를 조정한다. 여기서, 60X 1.42 개구 수 대물와 촛점 시스템을 사용하여 형광 이미지를 획득. </li></ol>정화조 마우스의 &quot;&gt; 4. 항생제 구조 <ol><li> 이전에 1 단계에 표시된대로 TIGR4 1.0 × 103 CFU 쥐에 감염 IP를 시작합니다. </li><li> 30 시간의 감염 후에서 시작하여, 36 시간 동안 100 ㎕의 생리 식염수에 12 시간마다 약제 학적 등급을 암피실린 IP 하였다 (80 mg / kg 체중)을 투여. 파라핀 삽입 및 조직 절편에 대한 단계 1.6에 설명 된대로 원하는 시점과 과정에서 마음을 수집합니다. </li></ol> 5. 콜라겐 증착 검출 <ol><li> 표준 방법 (17)를 사용하여 탈 이온수, deparaffinize 및 수화물 조직 섹션과 자일 렌 등급 알코올 세척을 사용. 5 분 동안 수성 0.2 % 인 몰리브덴 산으로 섹션을 처리하고, 5 분 동안 탈 이온수로 씻어냅니다. </li><li> 실온에서 2 시간 동안 피크르산 0.1 %와 시리우스 레드 염색 섹션. </li><li> 3 분 동안 0.01 N 염산 부분을 세척하고 1 분 동안 70 % 에탄올을 사용하여 씻어. </li><li> Dehydra동일한를 사용하여 섹션을 테하지만 5.1에 설명 된 것과 세척의 순서를 반대로, 자일 렌 증가에 심장 섹션을 탈수 한 후 에탄올의 농도를 등급. </li><li> 현미경 평가를위한 설치 솔루션 마운트 조직 섹션. </li><li> 콜라겐 침착의 영역을 식별하기 위해 광학 현미경을 사용하여 낮은과 높은 배율에서 깊이 읽기 염색 분야 스테인드 조직 섹션을 확인합니다. 빨간색 심장의 심실 내 모든 지역은 콜라겐이 증착 된 영역입니다. 이 붉은 얼룩없이 지역은 콜라겐 침착 부족 건강한 조직의 사이트입니다. 여기서, 0.5 20X 대물 개구를 구비 한 현미경을 사용 Picro 시리우스 레드 염색 이미지를 획득. 참고 : 건강한 조직에 빨간색 아트리움 얼룩. </li></ol>

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The authors declare no conflict of interest in this study.

이 보고서에서, 재현성이 높은 방법을 유도 S. 폐렴은 IPD 동안 생쥐의 심장 microlesions을 매개 성 및 시각화를위한 기술이 시연된다. 마우스는 인간 패혈증 동안 발생하고, 심근 궁극적으로 세균 전좌 리드 유사한 고급 균혈증, 선도, 클리어런스에 대한 면역 시스템의 임계치를 능가 박테리아의 투여로 감염된다. S., 아직 알 수없는 이유 마음에 폐렴는 호스트 면역 반응에 ...

지역 사회 획득 폐렴 (CAP)에 대한 성인의 입원 사망률 1-4에 기여 심장 관련 부작용에 대한 문서화 된 위험을 운반합니다. 코랄 레스 - 메디나 등의 등의 최근 연구에서. 심장 합병증과 및 / 또는 책임 폐렴 관련 사망 3의 27 %에 대해 관련이있는 것으로 밝혀졌다. 폐렴 구균 (폐렴), CAP 패혈증 5의 가장 흔한 원인, 직접 인정 성인 환자 6 많은 19 % 정도에서 심장 관련 부작용과 관련이있다. 폐렴과 관련된 심장 관련 부작용은 주파수 (6)의 내림차순 신규 또는 악화 울혈 성 심부전, 부정맥, 심근 경색 등이 있습니다. 브라운 등의 등의 최근 PLoS의 병원체 글에서., S. 폐렴은 myocardiu에 심장 혈관 내피 세포, 항목을 통해 전위 할 수있는 것으로 밝혀졌다m, 및 침습성 폐렴 구균 질환 (IPD) 7시 심실의 박테리아로 가득 이산 비 화농성 미세한 병변 (microlesions)의 형성. 심장 microlesion 형성의 증거는 폐렴 구균 감염에 굴복 한 인간이 아닌 영장류와 사람에서 심장 샘플에서 관찰되었다. 마찬가지로, 실험적으로 감염된 마우스는 재현성 심장 microlesions을 개발했다. 마우스에서 microlesion의 크기와 수는 긍정적 기간 및 균혈증, 혈청 심장 트로포 닌 수준의 심각도 및 비정상적인 심장 전기 생리학과의 상관 관계를 보였다. 마음에 세균 전좌는 혈액 - 뇌 장벽 및 폐렴 구균 성 수막염의 개발, 라미닌 수용체 혈소판, 혈관 내피 세포, 즉 콜린 결합 단백질 매개 내습 걸쳐 폐렴 전좌에 대한 책임 동일한 메커니즘을 통해 발생하는 것으로 밝혀졌다 활성화시키는 인자 수용체 의존적 8. Microles이온 형성은 심근 7 죽일 발견 된 폐렴 구균 기공 형성 독소 뉴 몰리이 필요합니다. 폐렴 구균 심장 microlesions은 황색 포도상 구균을 포함한 다른 그람 양성 세균에 의해 발생하는 화농성 연부 조직 및 심장 농양에서 구별된다. 이들은 호중구와 섬유소 침착 9,10 의해 둘러싸여 박테리아 단일 초점을 특징으로한다. Microlesions S.에 의해 형성 폐렴은 영향을받는 심장에 분포 크기, 작은, 그리고 면역 세포 침윤이 부족하다. 감염의 초기 단계 동안, microlesions는 S. 의한 폐렴은 심근 병증의 병적 징후를 연상 손상되거나 염증 조직의 영역으로 나타납니다. 일부 단핵구 그러나 그들의 존재는 수명이 짧고 병변은 빠른 속도로 동시에 GR 계속하면서 모양이 괴사되고 박테리아로 가득,이 시간 동안 관찰 할 수있다동물이나 항균 개입의 죽음까지 크기 아야. 중요한 항생제 개입 3 일 이내에, 넘치 면역 세포 침윤은 전 병변 부위에서 관찰이 강력한 콜라겐 침착을 수반한다. 유사 심장 리모델링은 심장 기능 11-15에 영향을 지속과 함께 경색에 따라 발생하는 것으로보고되었습니다. 따라서, microlesions 질병 에피소드에서 살아남은 회복기 개인의 심장 관련 사망의 발생률이 증가 IPD 동안 가능성이 발생하는 심장 관련 부작용에 ​​대한 잠재적 인 설명이다. 여기서, 명령은 감염의 초기와 후기 단계에서 IPD 심장 병변 형성 및 심장 microlesions의 가시화 실험 마우스 모델에 제공된다. 항균 개입하여 저장 한 동물에서 콜라겐 침착의 검출을위한 프로토콜이 설명된다. 이 문서의 목적은 F이다이 중요하고 새로운 폐렴 구균 병리에 다른 연구자의 연구를 acilitate.

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visualization of streptococcus pneumoniae within cardiac microlesions and subsequent cardiac remodeling

균혈증 동안 폐렴 구균은 심근에 혈관 내피 세포에서 이동시키다 및 이산 박테리아 채워진 미세한 병변으로 인해 면역 세포 침투의 부재로 주목받는 (microlesions)를 형성 할 수있다. 때문에 심독성 제품의 석방, S.에 microlesions 내 폐렴이 자주 성인의 뇌 산염 침습성 폐렴 구균 질환 동안 관찰되는 심부전에 기여하는 것으로 생각된다. 여기에서 30 시간 내에 심장 microlesion 형성을 재현하기 위해 리드 실험 마우스 감염에 대한 프로토콜을 설명된다. 명령이 헤 마톡 실린 및 스테인드 마음 섹션과 초기와 후기 microlesions의 형태 학적 차이를 에오신에 microlesion 식별에 제공된다 강조 표시됩니다. 명령은 S. 검증을위한 프로토콜에 제공되고 폐렴 구균 캡슐 다당류에 대한 항체를 사용하여 microlesions 내 폐렴면역 형광 현미경. 마지막으로, 감염된 마우스 및 검색 및 회복기 마우스의 마음의 흉터 형성의 평가를 구출 항생제 개입을위한 프로토콜을 제공한다. 함께, 이러한 프로토콜은 S. 노출의 결과로서 심근 침윤 폐렴, 심근 사망, 심근 개형 기본 분자 메커니즘의 연구를 촉진 할 것이다 폐렴과 심장의 폐렴 구균에 대한 면역 반응.

H &amp; E에 의해 Microlesions의 시각화 심장 microlesions 그림 1. TIGR4와 해당 IP의 반칙으로 IPD와 마우스의 심장 부분을 염색 H &amp; E의 심실에서 관찰 헤 마톡 실린의 더 많은 수의 24 및 30 시간 게시물 감염 (HPI) 사이의 이러한 병변의 크기의 증가를 보여줍니다했다 병변 내에서 볼 수 -stained (즉, 보라색 블루) 폐렴 구균. Microlesions는 액포 같은 형태, 밀도 에?...

Watch this Scientific Journal Video about Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling at JoVE.com

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals.

시각화<em&gt; 폐렴 구균</em&gt; 심장 Microlesions 및 후속 심장 리모델링 내

During bacteremia Streptococcus pneumoniae can translocate across the vascular endothelium into the myocardium and form discrete bacteria-filled ...

visualization-streptococcus-pneumoniae-within-cardiac-microlesions

Research

JoVE Journal

Medicine

Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

8.9K 조회수.  The University of Texas Health Science Center at San Antonio. Streptococcus pneumoniae forms discrete non-purulent microscopic lesions in the heart. Outlined is the protocol for a murine model of cardiac microlesion formation. Instruction is provided on microlesion visualization using microscopy, discrimination between early and late microlesions, and methods to detect cardiac remodeling in hearts of convalescent animals. 

비디오: Visualization of Streptococcus pneumoniae within Cardiac Microlesions and Subsequent Cardiac Remodeling

Cardiac Stress Test Induced by Dobutamine and Monitored by Cardiac Catheterization in Mice

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the arteries (&beta;2). When systemically administered, it increases cardiac demand. Thus, dobutamine unmasks abnormal rhythm or ischemic areas potentially at risk of infarction. 
Monitoring of heart function during a cardiac stress test can be performed by either ecocardiography or cardiac catheterization. The latter is an invasive but more accurate and informative technique that the former.
Cardiac stress test induced by dobutamine and monitored by cardiac catheterization accomplished as described here allows, in a single experiment, the measurement of the following hemodynamic parameters: heart rate (HR), systolic pressure, diastolic pressure, end-diastolic pressure, maximal positive pressure development (dP/dtmax) and maximal negative pressure development (dP/dtmin), at baseline conditions and under increasing doses of dobutamine.
As expected, in normal mice we observed a dobutamine dose-related increase in HR, dP/dtmax and dP/dtmin. Moreover, at the highest dose tested (12 ng/g/min) the cardiac decompensation of high fat diet-induced obese mice was unmasked.

We describe the protocol to perform a cardiac stress test induced by dobutamine and monitored by cardiac catheterization in normal mice. Also we show its application to unmask subclinical cardiac disease in high fat diet-induced obese mice.

심장 스트레스 테스트 Dobutamine에 의해 유도와 마우스의 심장 Catheterization에 의해 모니터링

Dobutamine is a &beta;-adrenergic agonist with an affinity higher for receptor expressed in the heart (&beta;1) than for receptors expressed in the ...

연구

의학

Direct Pressure Monitoring Accurately Predicts Pulmonary Vein Occlusion During Cryoballoon Ablation

Cryoballoon ablation (CBA) is an established therapy for atrial fibrillation (AF). Pulmonary vein (PV) occlusion is essential for achieving antral contact and PV isolation and is typically assessed by contrast injection. We present a novel method of direct pressure monitoring for assessment of PV occlusion.
Transcatheter pressure is monitored during balloon advancement to the PV antrum. Pressure is recorded via a single pressure transducer connected to the inner lumen of the cryoballoon. Pressure curve characteristics are used to assess occlusion in conjunction with fluoroscopic or intracardiac echocardiography (ICE) guidance. PV occlusion is confirmed when loss of typical left atrial (LA) pressure waveform is observed with recordings of PA pressure characteristics (no A wave and rapid V wave upstroke). Complete pulmonary vein occlusion as assessed with this technique has been confirmed with concurrent contrast utilization during the initial testing of the technique and has been shown to be highly accurate and readily reproducible.
We evaluated the efficacy of this novel technique in 35 patients. A total of 128 veins were assessed for occlusion with the cryoballoon utilizing the pressure monitoring technique; occlusive pressure was demonstrated in 113 veins with resultant successful pulmonary vein isolation in 111 veins (98.2%). Occlusion was confirmed with subsequent contrast injection during the initial ten procedures, after which contrast utilization was rapidly reduced or eliminated given the highly accurate identification of occlusive pressure waveform with limited initial training.
Verification of PV occlusive pressure during CBA is a novel approach to assessing effective PV occlusion and it accurately predicts electrical isolation. Utilization of this method results in significant decrease in fluoroscopy time and volume of contrast.

Effective pulmonary vein isolation utilizing a cryoballoon depends on complete pulmonary vein occlusion. The point of occlusion can be effectively predicted by direct analysis of pulmonary vein pressure waveform analysis during balloon inflation using a simple and reproducible technique.

직접 압력 모니터링 정확하게 Cryoballoon 절제하는 동안 폐 정맥 폐색을 예측

Cryoballoon  ablation (CBA) is an established therapy for atrial fibrillation (AF).  Pulmonary vein (PV) occlusion is essential for achieving antral ...

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and fluid retention. Changes in myocardial structure, electrical conduction, and energy metabolism develop with heart failure, leading to contractile dysfunction, increased risk of arrhythmias, and sudden death. Hypertensive heart disease is one of the key contributing factors of cardiac remodeling associated with heart failure. The most commonly-used animal model mimicking hypertensive heart disease is created via surgical interventions, such as by narrowing the aorta. Abdominal aortic constriction is a useful experimental technique to induce a pressure overload, which leads to heart failure. The surgery can be easily performed, without the need for chest opening or mechanical ventilation. Abdominal aortic constriction-induced cardiac pathology progresses gradually, making this model relevant to clinical hypertensive heart failure. Cardiac injury and remodeling can be observed 10 weeks after the surgery. The method described here provides a simple and effective approach to produce a hypertensive heart disease animal model that is suitable for studying disease mechanisms and for testing novel therapeutics.

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

쥐의 복부 대동맥의 수축을 통해 심장 리모델링의 모델

Heart failure is one of the leading causes of death worldwide. It is a complex clinical syndromethat includes fatigue, dyspnea, exercise intolerance, and ...

Preparation and In Vitro Characterization of Magnetized miR-modified Endothelial Cells

To date, the available surgical and pharmacological treatments for cardiovascular diseases (CVD) are limited and often palliative. At the same time, gene and cell therapies are highly promising alternative approaches for CVD treatment. However, the broad clinical application of gene therapy is greatly limited by the lack of suitable gene delivery systems. The development of appropriate gene delivery vectors can provide a solution to current challenges in cell therapy. In particular, existing drawbacks, such as limited efficiency and low cell retention in the injured organ, could be overcome by appropriate cell engineering (i.e., genetic) prior to transplantation. The presented protocol describes the efficient and safe transient modification of endothelial cells using a polyethyleneimine superparamagnetic magnetic nanoparticle (PEI/MNP)-based delivery vector. Also, the algorithm and methods for cell characterization are defined. The successful intracellular delivery of microRNA (miR) into human umbilical vein endothelial cells (HUVECs) has been achieved without affecting cell viability, functionality, or intercellular communication. Moreover, this approach was proven to cause a strong functional effect in introduced exogenous miR. Importantly, the application of this MNP-based vector ensures cell magnetization, with accompanying possibilities of magnetic targeting and non-invasive MRI tracing. This may provide a basis for magnetically guided, genetically engineered cell therapeutics that can be monitored non-invasively with MRI.

Preparation and In Vitro Characterization of Magnetized miR-modified Endothelial Cells

This manuscript describes the efficient, non-viral delivery of miR to endothelial cells by a PEI/MNP vector and their magnetization. Thus, in addition to genetic modification, this approach allows for magnetic cell guidance and MRI detectability. The technique can be used to improve the characteristics of therapeutic cell products.

준비 및<em&gt; 체외</em&gt; 자화 특성의 내피 세포를 미르 변성

To date, the available surgical and pharmacological treatments for cardiovascular diseases (CVD) are limited and often palliative. At the same time, gene ...

Cardiac Magnetic Resonance for the Evaluation of Suspected Cardiac Thrombus: Conventional and Emerging Techniques

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The appearance of a mass on certain magnetic resonance (MR) sequences can help differentiate a thrombus from competing diagnoses such as a tumor. T1 and T2 signal characteristics of a thrombus are related to the evolution of hemoglobin properties. A thrombus typically does not enhance following contrast administration, which also helps differentiation from a tumor. We also highlight the emerging role of T1 mapping in the evaluation of a thrombus, which can add another level of support in diagnosis. Prior to any CMR exam, patient screening and interviews are critical to ensure safety and to optimize patient comfort. Effective communication during the exam between the technologist and the patient promotes proper breath holding technique and higher quality images. Volumetric post processing and structured reporting are helpful to ensure that the radiologist answers the ordering services&#39; question and communicates these results effectively. Optimal pre-MR safety evaluation, CMR exam execution, and post exam processing and reporting allow for delivery of high quality radiological service in the evaluation of a suspected cardiac thrombus.

The goal of this article is to describe how cardiac magnetic resonance can be used for the evaluation and diagnosis of a suspected cardiac thrombus. The method presented will describe data acquisition as well as the pre-procedure and post-procedure protocol.

의심되는 심장 혈전의 평가를 위한 심장 자기 공명: 기존 및 신흥 기술

We present the conventional cardiac magnetic resonance (CMR) protocol for evaluating a suspected thrombus and highlight emerging techniques.&#160;The ...

Isolation and Purification of Murine Cardiac Pericytes

Pericytes, perivascular cells of microvessels and capillaries, are known to play a part in angiogenesis, vessel stabilization, and endothelial barrier integrity. However, their tissue-specific functions in the heart are not well understood. Moreover, there is currently no protocol utilizing readily accessible materials to isolate and purify pericytes of cardiac origin. Our protocol focuses on using the widely used mammalian model, the mouse, as our source of cells. Using the enzymatic digestion and mechanical dissociation of heart tissue, we obtained a crude cell mixture that was further purified by fluorescence activating cell sorting (FACS) by a plethora of markers. Because there is no single unequivocal marker for pericytes, we gated for cells that were CD31-CD34-CD45-CD140b+NG2+CD146+. Following purification, these primary cells were cultured and passaged multiple times without any changes in morphology and marker expression. With the ability to regularly obtain primary murine cardiac pericytes using our protocol, we hope to further understand the role of pericytes in cardiovascular physiology and their therapeutic potential.

We have optimized a protocol to isolate and purify murine cardiac pericytes for basic research and investigation of their biology and therapeutic potential.

뮤린 심장 pericytes의 격리 및 정화

Pericytes, perivascular cells of microvessels and capillaries, are known to play a part in angiogenesis, vessel stabilization, and endothelial barrier ...

Robotized Testing of Camera Positions to Determine Ideal Configuration for Stereo 3D Visualization of Open-Heart Surgery

Stereo 3D video from surgical procedures can be highly valuable for medical education and improve clinical communication. But access to the operating room and the surgical field is restricted. It is a sterile environment, and the physical space is crowded with surgical staff and technical equipment. In this setting, unobscured capture and realistic reproduction of the surgical procedures are difficult. This paper presents a method for rapid and reliable data collection of stereoscopic 3D videos at different camera baseline distances and distances of convergence. To collect test data with minimum interference during surgery, with high precision and repeatability, the cameras were attached to each hand of a dual-arm robot. The robot was ceiling-mounted in the operating room. It was programmed to perform a timed sequence of synchronized camera movements stepping through a range of test positions with baseline distance between 50-240 mm at incremental steps of 10 mm, and at two convergence distances of 1100 mm and 1400 mm. Surgery was paused to allow 40 consecutive 5-s video samples. A total of 10 surgical scenarios were recorded.

The human depth perception of 3D stereo videos depends on the camera separation, point of convergence, distance to, and familiarity of the object. This paper presents a robotized method for rapid and reliable test data collection during live open-heart surgery to determine the ideal camera configuration.

오픈-심장 수술의 스테레오 3D 시각화를 위한 이상적인 구성을 결정하기 위해 카메라 위치의 로봇화 테스트

Stereo 3D video from surgical procedures can be highly valuable for medical education and improve clinical communication. But access to the operating room ...

A Rodent Model of The Ross Operation: Syngeneic Pulmonary Artery Graft Implantation in A Systemic Position

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding root replacement, the possible dilation of the pulmonary autograft and subsequent aortic regurgitation is described. Several animal models have been proposed. However, these are usually limited to ex-vivo models or in-vivo experiments with relatively expensive large animal models. In this study, we sought to establish a rodent model of pulmonary artery graft (PAG) implantation in a systemic position. A total of 39 adult Lewis rats were included. Immediately after euthanasia, the pulmonary root was harvested from a donor animal (n=17). Syngeneic recipient (n=17) and sham-operated (n=5) rats were sedated and ventilated. In the recipient group, the PAG was implanted with an end-to-end anastomosis in infra-renal abdominal aortic position. Sham-operated rats underwent only transection and re-anastomosis of the aorta. Animals were followed with serial ultrasound studies for two months and post-mortem histological analysis. The median PAG diameter in the native position was 3.20 mm (IQR=3.18-3.23). At follow-up, the median diameter of the PAG was 4.03 mm (IQR=3.74-4.13) at 1 week, 4.07 mm (IQR=3.80-4.28) at 1 month, and 4.27 mm (IQR=3.90-4.35) at 2 months (p&lt;0.01). Peak systolic velocity was 220.07 mm/s (IQR=210.43-246.41) at 1 week, 430.88 mm/s (IQR=375.28-495.56) at 1 month, and 373.68 mm/s (IQR=305.78-429.81) at 2 months (p=0.02) and did not differ from the sham-operated group at the end of the experiment (p=0.5). Histological analysis did not show any sign of endothelial thrombosis. This study showed that rodent models may allow for the evaluation of the long-term adaptation of the pulmonary root to a high-pressure system. A systemically placed syngeneic PAG implantation represents a simple and feasible platform for the development and evaluation of novel surgical techniques and drug therapies to further improve the outcomes of the Ross operation.

We demonstrate how to establish a murine model of pulmonary root implantation into the descending aorta to simulate the Ross procedure. This model enables the medium/long-term evaluation of pulmonary autograft remodeling in a systemic position, representing the basis of developing therapeutic strategies to promote its adaptation.

로스 수술의 설치류 모델: 전신 위치에 있는 합성 폐 동맥 이식 이식

The Ross operation for aortic valve disease has regained new interest due to its outstanding long-term results. Nonetheless, when employed as freestanding ...

Large-Animal Model of Donation after Circulatory Death and Normothermic Regional Perfusion for Cardiac Assessment

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ donor pool. However, the DCD process is associated with the risk of cardiac tissue injury due to the inevitable period of warm ischemia. Normothermic regional perfusion (NRP) allows for an in situ organ assessment, allowing the procurement of hearts determined to be viable. Here, we describe a clinically relevant large-animal model of DCD followed by NRP. Circulatory death is established in anesthetized pigs by stopping mechanical ventilation. After a preset warm ischemia period, an extracorporeal membrane oxygenator (ECMO) is used for a NRP period lasting at least 30 min. During this reperfusion period, the model allows the collection of various myocardial biopsies and blood samples for initial cardiac evaluation. Once NRP is weaned, biochemical, hemodynamic, and echocardiographic assessments of cardiac function and metabolism can be performed before organ procurement. This protocol closely simulates the clinical scenario previously described for DCD and NRP in heart transplantation and has the potential to facilitate studies aimed at decreasing ischemia-reperfusion injury and enhance cardiac functional preservation and recovery.

The protocol describes a large-animal (porcine) model of donation after circulatory death, followed by thoracoabdominal normothermic regional perfusion that closely simulates the clinical scenario in heart transplantation, and has the potential to facilitate therapeutic studies and strategies.

순환기 사망 후 기증의 대형 동물 모델 및 심장 평가를 위한 정상 국소 관류

The increase in demand for cardiac transplantation throughout the years has fueled interest in donation after circulatory death (DCD) to expand the organ ...

Analysis of Cardiac Contractile Dysfunction and Ca2+ Transients in Rodent Myocytes

Contractile dysfunction and Ca2+ transients are often analyzed at the cellular level as part of a comprehensive assessment of cardiac-induced injury and/or remodeling. One approach for assessing these functional alterations utilizes unloaded shortening and Ca2+ transient analyses in primary adult cardiac myocytes. For this approach, adult myocytes are isolated by collagenase digestion, made Ca2+ tolerant, and then adhered to laminin-coated coverslips, followed by electrical pacing in serum-free media. The general protocol utilizes adult rat cardiac myocytes but can be readily adjusted for primary myocytes from other species. Functional alterations in myocytes from injured hearts can be compared to sham myocytes and/or to in vitro therapeutic treatments. The methodology includes the essential elements needed for myocyte pacing, along with the cell chamber and platform components. The detailed protocol for this approach incorporates the steps for measuring unloaded shortening by sarcomere length detection and cellular Ca2+ transients measured with the ratiometric indicator Fura-2 AM, as well as for raw data analysis.

Analysis of Cardiac Contractile Dysfunction and Ca2+ Transients in Rodent Myocytes

A set of protocols are presented that describe the measurement of contractile function via sarcomere length detection along with calcium (Ca2+) transient measurement in isolated rat myocytes. The application of this approach for studies in animal models of heart failure is also included.

설치류 근육 세포에서 심장 수축 기능 장애 및Ca 2+ 과도 현상 분석

Contractile dysfunction and Ca2+ transients are often analyzed at the cellular level as part of a comprehensive assessment of cardiac-induced injury ...

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