저자 홍 지앙 박사에게 감사를 표합니다 그리고 Deepa Parathasarthy, 시속, 기술 지원 BDS.

1. 전체 혈액 수집 <ol><li> 인간이나 NEM / EDTA (에틸렌 다이아 민 테트라 초산)가 들어있는 튜브에 실험 동물의 정맥 혈액을 수집합니다. </li><li> 즉시 7분 대한 rcf는 (상대 원심력) 혈장 및 적혈구 세포 펠렛을 준비하는 14,300에서 benchtop 원심에 혈액을 다운 스핀. </li><li> 고성능 액체 크로마 토그래피 (HPLC)와 chemiluminescence 검출 (CLD) 분석을 위해 플라즈마 샘플을 준비합니다. HPLC : 플라즈마, 석출물 플라즈마 단백질까지 10 분간 13,200 rpm으로 소용돌이와 원심 분리기에서 차가운 메탄올의 1시 1분 볼륨을 추가합니다. HPLC 분석을위한 뜨는 수집합니다. CLD : 나누어지는 구체적으로 분석 nitrosothiols에 sulfanilamide 및 mercuric 염화와 샘플 및 preinc​​ubate. </li><li> HPLC 및 CLD 분석에 붉은 세포 펠렛을 준비합니다. HPLC : 10 밀리미터 NEM, 2.5 밀리미터 EDTA (에틸렌 다이아 민 테트라 초산)과 ferricyanide 10 밀리미터를 포함 hypotonic 용해 용액에 추가 1시 4분 붉은 세포 펠릿. 와동은 철저하고 1시 1분 메탄올, 와동과 centrif 추가침전물 단백질까지 10 분 동안 13,200 rpm으로 uge. HPLC 분석을위한 뜨는 수집합니다. CLD. 10 밀리미터 EDTA (에틸렌 다이아 민 테트라 초산), 2.5 밀리미터 EDTA (에틸렌 다이아 민 테트라 초산)과 Ferricyanide 10 밀리미터를 포함 hypotonic 용해 용액에 1시 4분 붉은 세포 펠렛을 추가합니다. 나누어지는 샘플 nitrosothiols의 특정 검색에 대해 sulfanilamide 및 mercuric 염화물을 포함 튜브합니다. </li></ol> 2. 조직 추출 및 준비 <ol><li> NO metabolites의 조직 수준을 결정하기 위해서는 위에서 설명한 시료 준비를 위해 혈액을 무료로 조직을 수확 먼저 필요합니다. 전체 혈액의 교환은 좌심실의 꼭대기를 통해 생리 버퍼가 기적으로 열립니다. 모든 혈액이 제거되면 관심 휴지 그런 다음 수확 할 수 있습니다. </li><li> 조직 샘플을 Homogenize 및 HPLC 및 CLD 분석을 위해 위에서 설명한 샘플을 준비합니다. </li></ol> 3. 내피 기능에 대한 대동맥 반지 격리 조직 장기 목욕 <ol><li> 마우스가 anest 될 것입니다이상 반응 핀치를 상대하고, 이전 수술로 자궁 전위를 받아야 할 때까지 디에틸 에테르로 hetized. thoracotomy는 흉부 및 복부 대동맥을 폭로하기 위해 수행됩니다. 25 게이지 주사기는 좌심실의 꼭대기에 삽입 및 산소 Krebs Henseleit 버퍼와 혈액의 무료 perfused있다. </li><li> 오른쪽 심방은 혈액을위한 출구를 제공하기 위해 절단됩니다. 복부 대동맥이 제거되고 adventitia으로 지워지게됩니다. </li><li> 반지는 2mm 길이 세그먼트로 잘라 생리적 완충 용액으로 목욕 4 채널 조직 장기 목욕 (DMT 720MO, 광고 인 스트 루먼트)에 장착됩니다. </li><li> 선박 고리는 37 ° C.에 산소 Krebs 버퍼 (95% O 2 5 % CO 2)와 함께 10-ML 장기 욕조에 보관됩니다 한 g의 요구는 각 대동맥 고리 (이전 실험에서 결정된 최적 vasomotor 기능에 적합한 시작 장력)에 위치합니다. 여덟 채널 진수 브리지 (Powerlab) 및 데이터 수집 소프트웨어 (차트 버전 5.2.2)모든 힘을 측정을 기록하는 데 사용 기쁩니다. </li><li> 반지는 각 장기의 욕조에서 버퍼와 80분마다 20 분 동안 평형을 변경할 수있게됩니다. 80 분 대한 평형 후, 1 μm의 제품 phenylephrine은 submaximal 수축 각 고리에 추가됩니다. </li><li> 안정화되면, 이러한 아세틸콜린과 같은 내피 작용제는 선박 휴식 어떠한 생산과 노출된 정도를 확인하지에 추가됩니다. 아세틸콜린에 대한 선량 반응 후, 목욕탕은 질소 산화물, 평활근의 반응을 파악하고 100 %의 휴식을위한 가치를 얻을 나트륨 nitroprusside의 외인성 소스로 씻어서 recontracted 후 처리됩니다. </li><li> 어떠한 썩은 고기를 명확하게 선박 휴식 NO 및 억제의 석방을 설명하기 위해 욕조에 추가할 수 없습니다. </li></ol> 4. 대표 결과 이노-20 전용 HPLC의 이용은의 아질산염과 질산염의 구체적이고 민감한 검출을 위해 높은 처리량 방법을 사용하기 쉽게 제공합니다생물 학적 매트릭스. 감지 및 원래의 크로마토 그램의 원리는 그림 1에 표시됩니다. 이 방법은 아질산염과 질산염을 결정하는 생물 학적 샘플을 사용할 수 있습니다. NO metabolites의 CLD 기반 감지 NO의 분자 소스를 결정하는 화학 derivitization 단계가 필요합니다. chemiluminescence 검출기를위한 동시 산화 denitrosation 및 reductive denitrosation위한 실험 설정은 그림 2에 표시됩니다. 오른쪽의 반응 용기는 PBS 산도 7.4의 800mM ferricyanide으로 가득차 있으며, 왼쪽에있는 반응 용기는 감소 denitrosation위한 초산 (그림 2A)에 칼륨 요오드화물 / 요오드 혼합으로 가득합니다. 전체 설정은 그림 2B에 표시됩니다. 그림 3은 그룹의 특정 denitrosation의 아질산을 감지하고 계량 수 assays, nitrosothiols, nitrosamines뿐만 아니라 nitrosyl 헴 제품을 보여줍니다. 이 방법은 이전에 descri되었습니다침대 4,5 검증. 이러한 중요한 생화 학적 분석은 쉽게 내피 실험 동물에있어서 어떠한 생산을 결정하지 위해 절연 대동맥 고리에 대한 기능적 연구와 상호하실 수 있습니다. 이것은 고전 pharmacological 실험은 쉽고 정확하게 내피 기능과 생산 아니오를 평가할 수 없습니다. 이러한 아세틸콜린과 같은 내피 agonists에 선박 반응을 측정하는 것은 바로 다음 실험 동물의 혈액과 조직에서 발견 생화학 생체와 상호하실 수 있습니다 내피 어떠한 생산을 결정할 수 없습니다. 아세틸콜린에 대한 선량 반응의 전형적인 표현은 그림 4에 표시됩니다. 정상적인 내피 기능과 건강한 컨트롤 마우스는 휴식에 의해 아세틸콜린에 반응한다. 내피 기능 장애와 쥐 (hypercholesterolemic 생쥐)는 동일한 자극에 어떠한 생산을 줄일 수 없다 때문에 감소 휴식을 보여줍니다. <img alt="그림 1" src="/files/ftp_upload/3722/3722fig1.jpg" /> 무화과우레 1. 이노-20 및 샘플 크로마토 그램에 의한 아질산염과 질산염의 검출 원리. 아질산염과 질산염에 대한 검색의 이노-20 방식의 (위) 도식. (아래) 10 pmol의 아질산염과 질산염의 표준 chromotogram는 이노-20 (아질산염과 질산염 100 nm의 솔루션을 100 μl)에 주입. 100 μl 주입 부피 각 음이온 1 NM의 감도. 단백질이나 색 종 아무도 간섭 없습니다. <img alt="그림 2" src="/files/ftp_upload/3722/3722fig2.jpg" /> 정화 질소 산화물 가스의 기상 탐지와 요오드화물 / 요오드 검정을 사용 ferricyanide하고 reductive denitrosation에 의해 모두 산화 denitrosation를 사용하여 그림 2. CLD 실험 설정. <img alt="그림 3" src="/files/ftp_upload/3722/3722fig3.jpg" /> 그림 3. (패널) 그룹의 특정 chemic있는 샘플 preincubation 의해 reductive denitrosation 분석에서 아질산염의 Chemiluminescence 감지, RSNO, RNNO알 시약. 피크 영역의 뺄셈은 아질산염과 RSNOs의 검출을 허용합니다. ferricyanide의 산화 denitrosation 솔루션을 사용하여 nitrosyl 헴 종의 (패널 B) Chemiluminescent 탐지. 이 방법은 RSNOs (GSNO 또는 SNO-알부민) 또는 RNNO (NO-pyrrolidine와 N-nitroso-알부민)과 노 교차 반응으로 NO-헴 제품에만 적용됩니다. <img alt="그림 4" src="/files/ftp_upload/3722/3722fig4.jpg" /> 그림 4. 절연 대동맥 고리에서 전의 생체내 기관 목욕 후 HPLC 및 CLD에 의해 감지 생화학 생체와 상호하실 수 있습니다 내피 NO 생산의 직접적인 측정을 제공합니다. 이 수치는 더 이상 생산 감소하지 인해하도록 내피 기능 장애와 생쥐의 감소 휴식을 보여줍니다.

<ol>
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관심의 어떠한 충돌 선언 없습니다.

방법은 여러 생물 학적 구획의 관련 없음의 metabolites의 부량 위해 여기에서 설명한 내피에 의한 NO의 기능 측정과 상호 수있는 건강과 질병의 어떠한 생물의 지문 채취를 허용합니다. 이러한 방법은 높은 처리량을 위해 적응을위한 잠재력을 지닌 간단한 샘플 준비가 필요합니다. 이러한 분자의 상대적인 수량은 질환의 실험적 모델과 인간의 환자에서조차 직렬 샘플 수가 NO의 생산과 신진 대사 운?...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td></tr><tr><td> N-ethylmaleimide </td><td> 써모 과학 </td><td> 23,030 </td></tr><tr><td> EDTA (에틸렌 다이아 민 테트라 초산) </td><td> 시그마 - 올드 리치 </td><td> E7889 </td></tr><tr><td> Ferricyanide 칼륨 </td><td> Fluka </td><td> 60,299 </td></tr><tr><td> HPLC </td><td> Eicom 사 </td><td> 이노-20 </td></tr><tr><td> Autosampler </td><td> Alcott </td><td></td></tr><tr><td> DMT Myograph </td><td> 광고 인 스트 루먼트 </td><td></td></tr><tr><td> PowerLab </td><td> 광고 인 스트 루먼트 </td><td></td></tr><tr><td> Chemiluminescent </td><td> EcoPhysics </td><td> CLD 88Y </td></tr><tr><td> 원심 분리기 </td><td> Eppendorf </td><td> 5415D </td></tr><tr><td> 아세틸콜린 </td><td> 시그마 - 올드 리치</td><td> A6625 </td></tr><tr><td> R-(-) Phenylephrine </td><td> 시그마 - 올드 리치 </td><td> P6126 </td></tr></tbody></table>

analytical techniques for assaying nitric oxide bioactivity

질소 산화물은 (NO) 생물 학적 시스템 (혈액 순환에 이하 일초) 1로 극히 짧은 살고있다 이원 자유 라디칼이다. 아니오는 포함한 필수 기능을 조절, 우리의 신체에서 생산되는 가장 중요한 신호 분자 중 하나를 고려 없지만 혈압, 면역 반응 및 신경 통신 규제에 국한되지 수 있습니다. 따라서 생물 학적 매트릭스에서의 정확한 탐지 및 부량는 건강과 질병에서 NO의 역할을 이해하기 위해 중요합니다. NO 이러한 짧은 생리적 반 생활을 통해 어떠한 생화학의 반응 제품의 탐지를위한 대안 전략이 개발되었습니다. 여러 생물 학적 구획의 관련 없음의 metabolites의 양을 정함은 생체내에에 관해서는 어떠한 생산, 생체 이용률과 신진 대사에 중요한 정보를 제공합니다. 간단히 같은 혈액이나 혈장과 같은 하나의 구획을 샘플링하는 것은 전체 보의 정확한 평가를 제공하지 않을 수 있습니다특히 조직에서 어떠한 자격을 마구 없습니다. 실험 동물의 선택 조직으로 혈액을 비교하는 기능까지 건강과 질병의 어떠한 생체 진단 및 전조 유틸리티 등 기초 과학 및 임상 의학 간의 격차를 해소하는 데 도움이됩니다. 따라서 관심의 특정 조직에 혈장 또는 혈액 어떠한 상태의 추정은 더 이상 유효한 접근하지 않습니다. 그 결과, 방법 개발 및 생체내의 실험 동물의 여러 구획의 NO 및 NO-관련 제품 / metabolites의 탐지 및 부량을 허용하는 검증받을 것을 계속한다. 과 질산염 (NO 3 -) - 아질산 (NO 2) 최종 산화에 대한 수용성 guanylyl cyclase (SGC)의 활성화에 어떠한 synthases에 의해 생산에서 어떠한 생화학의 설립 패러다임은 생체내에서 NO의 효과의 일부분에 해당됩니다. S-nitrosothi을 형성하는 단백질 thiols, 차 아민 및 금속과 NO 및 NO-파생 metabolites의 상호 작용ols (RSNOs), N-nitrosamines (RNNOs) 및 nitrosyl-헴는 각각 NO의 cGMP 독립적인 효과를 나타내며 어떠한 의한 SGC의 활성화로 생리학 가능성만큼이나 중요하다. 생리학에서 NO의 진정한 이해는 동시에 여러 구획을 샘플링 생체내 실험에서에서 파생됩니다. 질소 산화물 (NO) 방법론은 어떠한 생화학에 대해서도 많은 논쟁과 토론의 복잡하고 종종 혼란 과학 및 초점이다. 우리의 능력을 구체적에 대한 어떠한 경첩과 관련된없는 새로운 메커니즘 및 신호 전달 경로의 해설은 선택과 sensitively 눈치채지 못한 어떠한와 관련된 모든 복잡한 생물 학적 매트릭스의 어떠한 제품과 metabolites을 수량. 여기서는 HPLC에 의한 아질산염과 질산염의 신속하고 민감한 분석 방법뿐만 아니라 NO의 분자 소스뿐만 아니라 전 생체내과를 결정하는 화학 derivitazation와 체외의 오존 기반 chemiluminescence에서 사용하는 생물 학적 시료의 여유 없음의 검출을 제시장기 목욕 myography.

Watch this Scientific Journal Video about Analytical Techniques for Assaying Nitric Oxide Bioactivity at JoVE.com

Analytical Techniques for Assaying Nitric Oxide Bioactivity

질소 산화물 (NO)의 내생적인 생산은 생물 학적 기능의 다양한 조절. 그것은 어떠한 기반 시그널링의 장애 또는 dysregulation이 많은 사람이 질병에 연루되어 있는지 점점 더 분명 해지고 있습니다. 메소드는 더 이상 metabolites 인간 질병에 대한 소설 진단 또는 전조 생체 자료를 제공할 수 없습니다 관련된 계량합니다.

질소 산화물 Bioactivity을 시금을위한 분석 기법

Nitric oxide (NO) is a diatomic free radical that is extremely short lived in biological systems (less than 1 second in circulating blood)1. NO may be ...

analytical-techniques-for-assaying-nitric-oxide-bioactivity

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JoVE Journal

Medicine

17.9K 조회수.  University of Texas Health Science Center at Houston. 질소 산화물 (NO)의 내생적인 생산은 생물 학적 기능의 다양한 조절. 그것은 어떠한 기반 시그널링의 장애 또는 dysregulation이 많은 사람이 질병에 연루되어 있는지 점점 더 분명 해지고 있습니다. 메소드는 더 이상 metabolites 인간 질병에 대한 소설 진단 또는 전조 생체 자료를 제공할 수 없습니다 관련된 계량합니다.

비디오: Analytical Techniques for Assaying Nitric Oxide Bioactivity

5/6th Nephrectomy in Combination with High Salt Diet and Nitric Oxide Synthase Inhibition to Induce Chronic Kidney Disease in the Lewis Rat

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements of homeostasis, integrative animal models are necessary to study development and progression of CKD. To study development of CKD and novel therapeutic interventions in CKD, we use the 5/6th nephrectomy ablation model, a well known experimental model of progressive renal disease, resembling several aspects of human CKD. The gross reduction in renal mass causes progressive glomerular and tubulo-interstitial injury, loss of remnant nephrons and development of systemic and glomerular hypertension. It is also associated with progressive intrarenal capillary loss, inflammation and glomerulosclerosis. Risk factors for CKD invariably impact on endothelial function. To mimic this, we combine removal of 5/6th of renal mass with nitric oxide (NO) depletion and a high salt diet. After arrival and acclimatization, animals receive a NO synthase inhibitor (NG-nitro-L-Arginine) (L-NNA) supplemented to drinking water (20 mg/L) for a period of 4 weeks, followed by right sided uninephrectomy. One week later, a subtotal nephrectomy (SNX) is performed on the left side. After SNX, animals are allowed to recover for two days followed by LNNA in drinking water (20 mg/L) for a further period of 4 weeks. A high salt diet (6%), supplemented in ground chow (see time line Figure 1), is continued throughout the experiment. Progression of renal failure is followed over time by measuring plasma urea, systolic blood pressure and proteinuria. By six weeks after SNX, renal failure has developed. Renal function is measured using 'gold standard' inulin and para-amino hippuric acid (PAH) clearance technology. This model of CKD is characterized by a reduction in glomerular filtration rate (GFR) and effective renal plasma flow (ERPF), hypertension (systolic blood pressure&gt;150 mmHg), proteinuria (&gt; 50 mg/24 hr) and mild uremia (&gt;10 mM). Histological features include tubulo-interstitial damage reflected by inflammation, tubular atrophy and fibrosis and focal glomerulosclerosis leading to massive reduction of healthy glomeruli within the remnant population (&lt;10%). Follow-up until 12 weeks after SNX shows further progression of CKD.

A two-stage method to establish chronic kidney disease (CKD) in the Lewis rat by surgically removing 5/6th of renal mass is described. Combination of the surgical procedure, NOS-inhibition and a high-salt diet leads to a model resembling human CKD, allowing study of causal mechanisms and development of novel therapeutic interventions.

높은 소금 다이어트와 루이스 쥐에서 만성 신장 질환을 유발하는 산화 질소 합성 효소 억제와 함께 5/6th의 절제

Chronic kidney disease (CKD) is a global problem. Slowing CKD progression is a major health priority. Since CKD is characterized by complex derangements ...

연구

의학

Techniques for Processing Eyes Implanted With a Retinal Prosthesis for Localized Histopathological Analysis

With the recent development of retinal prostheses, it is important to develop reliable techniques for assessing the safety of these devices in preclinical studies. However, the standard fixation, preparation, and automated histology procedures are not ideal. Here we describe new procedures for evaluating the health of the retina directly adjacent to an implant. Retinal prostheses feature electrode arrays in contact with eye tissue. Previous methods have not been able to spatially localize the ocular tissue adjacent to individual electrodes within the array. In addition, standard histological processing often results in gross artifactual detachment of the retinal layers when assessing implanted eyes. Consequently, it has been difficult to assess localized damage, if present, caused by implantation and stimulation of an implanted electrode array. Therefore, we developed a method for identifying and localizing the ocular tissue adjacent to implanted electrodes using a (color-coded) dye marking scheme, and we modified an eye fixation technique to minimize artifactual retinal detachment. This method also rendered the sclera translucent, enabling localization of individual electrodes and specific parts of an implant. Finally, we used a matched control to increase the power of the histopathological assessments. In summary, this method enables reliable and efficient discrimination and assessment of the retinal cytoarchitecture in an implanted eye.

Techniques for visualizing retinal cytoarchitecture directly adjacent to individual electrodes within a retinal stimulator.

현지화 된 조직 병리학 적 분석을 위해 망막 보철물 이식 처리 눈을위한 기법

With  the recent development of retinal prostheses, it is important to develop  reliable techniques for assessing the safety of these devices in ...

Experimental and Imaging Techniques for Examining Fibrin Clot Structures in Normal and Diseased States

Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on fibrin in the past years to include the investigation of synthesis, structure-function, and lysis of clots. However, there is still much unknown about the morphological and structural features of clots that ensue from patients with disease. In this research study, experimental techniques are presented that allow for the examination of morphological differences of abnormal clot structures due to diseased states such as diabetes and sickle cell anemia. Our study focuses on the preparation and evaluation of fibrin clots in order to assess morphological differences using various experimental assays and confocal microscopy. In addition, a method is also described that allows for continuous, real-time calculation of lysis rates in fibrin clots. The techniques described herein are important for researchers and clinicians seeking to elucidate comorbid thrombotic pathologies such as myocardial infarctions, ischemic heart disease, and strokes in patients with diabetes or sickle cell disease.

In this manuscript, experimental techniques, including blood preparation, confocal microscopy, and lysis rate analysis, to examine the morphological differences between normal and abnormal clot structures due to diseased states are presented.

Fibrin is an extracellular matrix protein that is responsible for maintaining the structural integrity of blood clots. Much research has been done on ...

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic study in the liver. Here, numerous assays are described to comprehensively measure the phenotype and parameters of hepatic steatosis in mouse and hepatocyte models. 
Combining the physiological, histological, and biochemical methods, this system can be used to assess the progress of hepatic steatosis. In vivo, the measurements of body weight and nuclear magnetic resonance (NMR) provide a general understanding of mice in a non-invasive manner. Hematoxylin and Eosin (H&amp;E) and Oil Red O staining determine the histological morphology and lipid deposition of liver tissue under nutrient overload conditions, such as high-fat diet feeding. Next, the total lipid contents are isolated by chloroform/methanol extraction, which are followed by a biochemical analysis for triglyceride and cholesterol. Moreover, mouse primary hepatocytes are treated with high glucose plus insulin to stimulate lipid accumulation, an efficient in vitro model to mimic diet-induced hyperglycemia and hyperinsulinemia in vivo. Then, the lipid deposition is measured by Oil Red O staining and chloroform/methanol extraction. Oil Red O staining determines intuitive hepatic steatotic phenotypes, while the lipid extraction analysis determines the parameters that can be analyzed statistically. The present protocols are of interest to scientists in the fields of fatty liver diseases, insulin resistance, and type 2 diabetes.

Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis

Here, optimized methods to generate in vivo and in vitro models of hepatic steatosis and to analyze the steatotic phenotypes and related physiological parameters are described.

의 최적화 분석<em&gt; 생체</em&gt; 및<em&gt; 체외</em&gt; 간 지방증

Establishing a system of procedures to qualitatively and quantitatively characterize in vivo and in vitro hepatic steatosis is important for metabolic ...

Synthesis, Characterization, and Application of Superparamagnetic Iron Oxide Nanoprobes for Extrapulmonary Tuberculosis Detection

A molecular imaging probe comprising superparamagnetic iron oxide (SPIO) nanoparticles and Mycobacterium tuberculosis surface antibody (MtbsAb) was synthesized to enhance imaging sensitivity for extrapulmonary tuberculosis (ETB). An SPIO nanoprobe was synthesized and conjugated with MtbsAb. The purified SPIO-MtbsAb nanoprobe was characterized using TEM and NMR. To determine the targeting ability of the probe, SPIO-MtbsAb nanoprobes were incubated with Mtb for in vitro imaging assays and injected into Mtb-inoculated mice for in vivo investigation with magnetic resonance (MR). The contrast enhancement reduction on magnetic resonance imaging (MRI) of Mtb and THP1 cells showed proportional to the SPIO-MtbsAb nanoprobe concentration. After 30 min of intravenous SPIO-MtbsAb nanoprobe injection into Mtb-infected mice, the signal intensity of the granulomatous site was enhanced by 14-fold in the T2-weighted MR images compared with that in mice receiving PBS injection. The MtbsAb nanoprobes can be used as a novel modality for ETB detection.

To improve serological diagnostic tests for Mycobacterium tuberculosis antigens, we developed superparamagnetic iron oxide nanoprobes to detect extrapulmonary tuberculosis.

초초파라마그네틱 산화질소 나노프로브의 합성, 특성화 및 적용

A molecular imaging probe comprising superparamagnetic iron oxide (SPIO) nanoparticles and Mycobacterium tuberculosis surface antibody (MtbsAb) was ...

Platelet-based Detection of Nitric Oxide in Blood by Measuring VASP Phosphorylation

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent vasoactive agents, nitric oxide (NO), can also act as a powerful inhibitor of platelet aggregation. Direct NO detection in blood is very challenging due to its high reactivity with cell-free hemoglobin that limits NO half-life to the millisecond range. Currently, NO changes after interventions are only estimated based on measured changes of nitrite and nitrate (members of the nitrate-nitrite-NO metabolic pathway). However precise, these measurements are rather difficult to interpret vis a vis actual NO changes, due to the naturally high baseline nitrite and nitrate levels that are several orders of magnitude higher than expected changes of NO itself. Therefore, the development of direct and simple methods that would allow one to detect NO directly is long overdue. This protocol addresses a potential use of platelets as a highly sensitive NO sensor in blood. It describes initial platelet rich plasma (PRP) and washed platelet preparations and the use of nitrite and deoxygenated red blood cells as NO generators. Phosphorylation of VASP at serine 239 (P-VASPSer239) is used to detect the presence of NO. The fact that VASP protein is highly expressed in platelets and that it is rapidly phosphorylated when NO is present leads to a unique opportunity to use this pathway to directly detect NO presence in blood.

Here, we present a protocol to address the potential use of platelets as a highly sensitive nitric oxide sensor in blood. It describes initial platelet preparation and the use of nitrite and deoxygenated red blood cells as nitric oxide generators.

VASP 인 산화를 측정 하 여 혈액에 산화 질소의 혈소판 기반 탐지

Platelets are the blood components responsible for proper blood clotting. Their function is highly regulated by various pathways. One of the most potent ...

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 ...

Tracking Superparamagnetic Iron Oxide-labeled Mesenchymal Stem Cells using MRI after Intranasal Delivery in a Traumatic Brain Injury Murine Model

Stem cell-based therapies for brain injuries, such as traumatic brain injury (TBI), are a promising approach for clinical trials. However, technical hurdles such as invasive cell delivery and tracking with low transplantation efficiency remain challenges in translational stem-based therapy. This article describes an emerging technique for stem cell labeling and tracking based on the labeling of the mesenchymal stem cells (MSCs) with superparamagnetic iron oxide (SPIO)&#160;nanoparticles, as well as intranasal delivery of the labeled MSCs. These nanoparticles are fluorescein isothiocyanate (FITC)-embedded and safe to label the MSCs, which are subsequently delivered to the brains of TBI-induced mice by the intranasal route. They are then tracked non-invasively in vivo by real-time magnetic resonance imaging (MRI). Important advantages of this technique that combines SPIO for cell labeling and intranasal delivery include (1) non-invasive, in vivo MSC tracking after delivery for long tracking periods, (2) the possibility of multiple dosing regimens due to the non-invasive route of MSC delivery, and (3) possible applications to humans, owing to the safety of SPIO, non-invasive nature of the cell-tracking method by MRI, and route of administration.

Presented here is a protocol for non-invasive mesenchymal stem cell (MSC) delivery and tracking in a mouse model of traumatic brain injury. Superparamagnetic iron oxide&#160;nanoparticles are employed as a magnetic resonance imaging (MRI) probe for MSC labeling and non-invasive in vivo tracking following intranasal delivery using real-time MRI.

외상성 뇌 손상 뮤린 모델에서 비강 내 전달 후 MRI를 사용하여 수퍼 파라마그네틱 철 산화물 표지 메젠키말 줄기 세포 추적

Stem cell-based therapies for brain injuries, such as traumatic brain injury (TBI), are a promising approach for clinical trials. However, technical ...

Near Infrared Photoimmunotherapy for Mouse Models of Pleural Dissemination

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination model can be used for the evaluation of treatment methods for intrathoracic diseases such as lung cancer or malignant pleural mesothelioma.
Near-infrared photoimmunotherapy (NIR-PIT) is a recently developed cancer treatment strategy that combines the specificity of tumor-targeting antibodies with toxicity caused by a photoabsorber (IR700Dye) after exposure to NIR light. The efficacy of NIR-PIT has been reported using various antibodies; however, only a few reports have shown the therapeutic effect of this strategy in an orthotopic model. In the present study, we demonstrate an example of efficacy evaluation of the pleural disseminated lung cancer model, which was treated using NIR-PIT.

Near-infrared photoimmunotherapy (NIR-PIT) is an emerging cancer therapeutic strategy that utilizes an antibody-photoabsorber (IR700Dye) conjugate and NIR light to destroy cancer cells. Here, we present a method to evaluate the antitumor effect of NIR-PIT in a mouse model of pleural disseminated lung cancer and malignant pleural mesothelioma using bioluminescence imaging.

흉막 보급의 마우스 모형을 위한 근적외선 광면역요법

The efficacy of photoimmunotherapy can be evaluated more accurately with an orthotopic mouse model than with a subcutaneous one. A pleural dissemination ...

A Novel Inhalation Mask System to Deliver High Concentrations of Nitric Oxide Gas in Spontaneously Breathing Subjects

Nitric Oxide (NO) is administered as gas for inhalation to induce selective pulmonary vasodilation. It is a safe therapy, with few potential risks even if administered at high concentration. Inhaled NO gas is routinely used to increase systemic oxygenation in different disease conditions. The administration of high concentrations of NO also exerts a virucidal effect in vitro. Owing to its favorable pharmacodynamic and safety profiles, the familiarity in its use by critical care providers, and the potential for a direct virucidal effect, NO is clinically used in patients with coronavirus disease-2019 (COVID-19). Nevertheless, no device is currently available to easily administer inhaled NO at concentrations higher than 80 parts per million (ppm) at various inspired oxygen fractions, without the need for dedicated, heavy, and costly equipment. The development of a reliable, safe, inexpensive, lightweight, and ventilator-free solution is crucial, particularly for the early treatment of non-intubated patients outside of the intensive care unit (ICU) and in a limited-resource scenario. To overcome such a barrier, a simple system for the non-invasive NO gas administration up to 250 ppm was developed using standard consumables and a scavenging chamber. The method has been proven safe and reliable in delivering a specified NO concentration while limiting nitrogen dioxide levels. This paper aims to provide clinicians and researchers with the necessary information on how to assemble or adapt such a system for research purposes or clinical use in COVID-19 or other diseases in which NO administration might be beneficial.

This simple and highly adaptable system device for the inhalation of high-concentration nitric oxide (NO) gas does not require mechanical ventilators, positive pressure, or high gas flows. Standard medical consumables and a snug-fitting mask are used to safely deliver NO gas to spontaneously breathing subjects.

자연적으로 호흡하는 과목에서 고농도의 산화산화물 가스를 전달하는 새로운 흡입 마스크 시스템

Nitric Oxide (NO) is administered as gas for inhalation to induce selective pulmonary vasodilation. It is a safe therapy, with few potential risks even if ...