이 연구는 독일 연구 협회 (CRC1039 A3) 및 연구 자금 지원 프로그램 헤센, 중개 의학 및 약학 TMP 연구 센터의 주 &quot;Landesoffensive 주르 ENTWICKLUNG wissenschaftlich-ökonomischer Exzellenz&quot;(LOEWE)과 그렇지 크로네-는 Fresenius 재단 지원 (EKFS), 연구 교육 그룹 중개 연구 혁신 - 제약 (TRIP).

SJL 1. EAE 유도 / J 마우스 <ol><li> 마우스 <ol><li> 11 주 된 여성 SJL / J 마우스를 사용하고이 약 7 일 동안 실험 방으로 길들 할 수 있습니다. 그룹 당 N = 10 마우스를 사용합니다. </li><li> 약물 효과의 평가를 위해 연속적으로 식수 통하거나 3 오일 면역화 후 (N = 그룹당 10)을 출발 음식 펠릿 통해 대조군 약물 위약을 투여. 질병의 피크 동안 우유 또는 3 % 설탕 물에 젖은 콘플레이크와 약물 또는 위약을 관리 할 수 ​​있습니다. </li></ol></li><li> 면역 물질 <ol><li> 완전한 프로 인트 보조제 (CFA, 열 사망 결핵균 H37 Ra)가 2 병 (5 μg의 각)와 에멀젼에 EAE 유도 키트 항원으로 구성된 (테오 단백질의 펩티드, PLP139-151, 1 ㎎ / ㎖ 에멀젼)를 사용하여 동결 건조 백일해 독소 (PTX). </li><li> 1X 인산염 완충 식염수에서 PTX (2 μg의 / ㎖) (PBS를 녹이고, 즉, &lt;/ EM&gt;에서) 같은 피펫 팁과 제거, 잘 혼합하고, 50 mL의 튜브에 PBS 1 ㎖에 추가하는 각 PTX 튜브에 PBS 1.5 mL를 넣어; 잘 섞다. </li></ol></li><li> 면제 <ol><li> 꼬리의 기부에,이 부분의 각 100 μL를 모두 PLP / CFA 피하 주입한다. 상단 뒤쪽이나 목의 피부에있는 면역 반응이 머리와 척수의 영상을 방해하기 때문에, 목에의 뒷면에 삽입하지 마십시오. </li><li> 접종 후 2 시간 및 24 시간의 초 - 당 두번 마우스 복강 PTX (IP) 100 μL, 상기 제 1 주입한다. </li><li> 제어 생쥐 들어 PLP없이 PLP없이 CFA (100 μL의 두 부분) 플러스 PTX 주입. </li></ol></li><li> 마우스 예방 접종 후 처리 <ol><li> 쥐에게 하루 7 매일 최대의 무게를 측정 한 다음 매일을 무게. 주 : 마우스 1 잃게 - EAE 동안 체중을 2g. 감소는 EAE의 발병을 표시합니다. </li><li> 7 일 협정 매일 임상 증상을 평가운동 기능의 명백한 변화를 0.5 점수 : 표준 점수 시스템에 보내고 (즉, 0 점수 없습니다 꼬리의 말단 마비 1 점수 : 전체 꼬리 마비를 1.5 점수 : 하나 또는 두 뒷다리의 가벼운 마비 2 점수 : 뒷 다리의 심한 마비, 점수 2.5 : 하나의 뒷 다리의 완전 마비; 골 3 : 두 뒷다리의 완전한 마비, 3.5 점수 :. 뒷다리 하나 앞 다리의 마비의 완전 마비는 3.5 이상의 점수에 대한 함께 쥐를 안락사 &gt; 12 시간. </li></ol></li><li> EAE 코스와 영상의 시간 <ol><li> 접종 후 12 - 마우스는 10 일째 주위 발생 점수&gt; 1에 도달하면,이 질병의 발병에 제 1 이미징을 수행한다. </li><li> 삼일 - 초기 증상이 후 1 ~ 2 일에 도달되며, 1 동안 지속됩니다 피크에서 두 번째 영상을 수행합니다. 참고 : 그 후, 쥐가 완전히 7 ~ 10 일 이내에 복구합니다. 간격 동안의 이미지는 여전히 혈관 누수를 표시 할 수 있습니다하지만 염증 지표는 음수가 될 것이다. </li></ol></li></ol> 2. 발광 생물과 시신경염 및 뇌 염증의 근적외선 영상 <ol><li> 촬상 시스템의 설치 <ol><li> 생물 발광 근적외선 신호의 분석을 가능하게하는 장치와 생체 내 영상에 수행한다. </li><li> 모든 이미징 과정에서 2.5 %의 이소 플루 란 마취 - 2에서 쥐를 유지합니다. </li><li> 위치 하나 중간에 가스 공급 장치를 사용하여 장치에서 서로 옆에 두 개의 마우스. 중앙의 상부 척추를 놓습니다. </li><li> 약물 치료 효과의 평가가 쌍을 비교하기 위해, 동시에 두 개의 마우스, 그룹 당 하나를 사용하십시오. 이 생물 발광 이미징을위한 중요합니다. </li><li> 검은 천으로 예방 접종의 사이트를 보호하고 마우스 / 마우스의 정확한 위치를 평가하기 위해 사진과베이스 라인 이미지를 촬영합니다. 모든 이미지를 카메라에 6.5 cm의 거리를두고 B-초점을 사용합니다. </li></ol></li><li> 주입 및 생물 발광 염증 프로브 이미징 <ol><li> 생체 내 이미징 시스템 설정을 사용 : 8 비닝 에피 BLI는, 엠가 열려 필터링, 전 필터 블록, FSTOP 1, B = 6.5 cm, 광고 노출 (120)의 초점; 베이스 라인 이미지를 촬영. </li><li> 즉시 사용 가능한 화학 발광 시약 (40 ㎎ / ㎖)의 100 μL의 IP를 주입한다. 주사기를 채우기 전에 잘 섞는다. </li><li> 생물 발광 이미지를 5, 10, 및 주입 후 15 분 캡처. 생물 발광 피크의 시간 경과 동물간에 다르다. 주 : 최대 5 발생합니다 - 주입 후 10 분; 15 분에서 감소는 더 이상 이미지가 필요하지 않습니다 나타냅니다. 때문에 서로 다른 시간의 과정에 약간의 편견을 제거하기 위해 제어 및 치료 그룹의 마우스 쌍을 사용합니다. </li><li> 실험에 관련된 설명을 입력합니다. 자동 팝업 대화 상자를 관찰; 이 마우스 변형, 성별, 시점, 프로브 주입, 그룹 등의 시간 등의 정보를 포함한다. 파일 모두 O를 저장북동 폴더; 그들은 시간 태그와 모든 설명을해야합니다. </li></ol></li><li> BBB 무결성을위한 근적외선 형광 나노 입자의 주입 및 이미징 <ol><li> B 조 초점 (6.5 cm 거리)에 근적외선 표면 형광 이미징을 사용합니다. 페 길화 된 형광 나노 입자의 여기 / 발광 최대 값은 690분의 675 나노 미터이다. Ex640 / Em700 및 Ex675 / Em720에서 서로 다른 파장, 두 개의 이미지를 캡처; 8 비닝 2의 노출을 사용하고 FSTOP 2.베이스 라인 이미지를 가져 가라. </li><li> 꼬리 정맥을 통해 페 길화 된 형광 적외선 나노 입자의 70 μL 정맥 주사하고 쥐 3 시간과 위의 설정을 사용하여 주사 후 24 시간을 상상한다. 주사기를 채우기 전에 용액을 잘 섞는다. </li><li> 신호의 특이성을 평가하기 위해 필요하다 대조군 마우스에 0.9 % 염화 나트륨 주입한다. </li></ol></li><li> 주입 및 MMP 활성을위한 근 적외 형광 프로브의 이미징 <ol><li> 면도를하거나 머리를 털을 뽑다 및 유베이스 라인 이미지를 복용하기 전에 조심스럽게 일일 pper 척추 영역입니다. 피부가 손상되지 않아야합니다. </li><li> B 조 초점 (6.5 cm 거리)에 근적외선 표면 형광 이미징을 사용합니다. MMP를 기동 프로브의 여기 / 발광 최대 값은 700분의 680 나노 미터이다. Ex640 / Em700 및 Ex675 / Em720에서 서로 다른 파장, 두 개의 이미지를 캡처; 8 비닝 1의 노출을 사용하고 FSTOP 2.베이스 라인 이미지를 가져 가라. </li><li> 이 마우스 (10)에 충분하게되도록 즉시 사용 가능한 용액 (1X PBS 20을 nmol / 1.5 mL)을 1.5 ㎖의 튜브에 200 μL의 PBS 1X에 추가; 주사기를 채우기 전에 잘 섞는다. 참고 : 제공된 볼륨이 약간의 볼륨이 주사기 충전 및 주입시 손실을 고려하지 않습니다. </li><li> 촬상 전에 꼬리 정맥을 통해 24 시간 프로브 IV 150 μL를 주입한다. 신호의 특성을 평가하는 경우에만 대조군으로 PBS를 주입한다. </li><li> 주사 후 24 시간에서, 적어도 2 개의 파장, Ex640 / Em700 및 예 675 / E에 에피 FL 이미지를 가지고m720은 설정 (8 및 FSTOP 2 비닝 1의 노출, 초점 B) 위 설명했다. 주의 : 2 개의 파장의 사용이 비특이적 신호를 감산하여 스펙트럼 unmixing 허용한다. </li></ol></li></ol> 3. 이미지 분석 <ol><li> 생물 발광 분석 (BLI) <ol><li> 를 열 수있는 소프트웨어를 두 번 클릭합니다. </li><li> 상단 메뉴 바에서 실험의 폴더의 디렉토리로 이동을 선택, 파일 브라우저 아이콘을 클릭; 이 테이블에있는 폴더의 모든 파일을 엽니 다. </li><li> 실험에 관련된 설명을 표시하는 열을 구성합니다. </li><li> 품질 관리의 경우, EAE 신호의 특이성을 확인 EAE 및 / 또는 순진한 마우스의 증상없이 비 응답자 마우스의 파일을 더블 클릭합니다. 참고 : 순진 마우스와 비 응답자 마우스에서 최소한의 신호에는 신호가 없어야합니다. <ol><li> 프로의 주입하기 전에 기본 이미지를 확인신호의 특이성을 추가 대조군으로서 각 마우스 일; 그것은 부정적인해야합니다. </li></ol></li><li> 이미지를 선택하려면 첫 번째 EAE 마우스의 첫 번째 파일을 더블 클릭하고 생물 발광 강도 (LUT 바 범위) 및 현지화를 확인합니다. 모든 이미지를 하나씩 확인합니다. 각 마우스의 낮은 강도로 파일을 닫습니다 (즉, 각 마우스 2 3 중 이미지를 유지). </li><li> 이미지 조정 및 수출 <ol><li> 정량화 할 수있는 첫 번째 파일을 두 번 클릭합니다. 새 창 팝 업을 준수하십시오. &quot;옵션&quot;(상위 메뉴)에서 각 이미지에 표시 할 라벨을 사용자 정의 할 수 있습니다. </li><li> 오른쪽 도구 팔레트에서 이동 &quot;이미지 조정.&quot; 기본적으로 최소 및 최대 강도는 &quot;자동&quot;으로 설정하고 무지개 사색에 표시됩니다. 필요한 경우 설정을 변경하려면 &quot;사용 설명서&quot;를 선택합니다. 참고 : 예를 들어, 모든 내 보낸 이미지가 동일한 LUT 바 (동일 최소값과 최대 값)을 할 수 있습니다 쉽게 비교할 수 있습니다.최소값과 최대 값의 조정은 양적 결과에 영향을주지 않습니다. </li><li> 이미지 내보내기를 선택 PNG, 디렉토리 및 이미지 이름을 클릭 </li></ol></li><li> 관심 지역의 정량 (ROI) <ol><li> 도구 팔레트에서 &quot;ROI&quot;도구로 이동합니다. 투자 수익 (ROI) 방법 (원형, 사각형, 자동차, 또는 자유 손)와 로아의 수를 선택합니다. ROI를 창 이미지 창 팝업 관찰한다. </li><li> 마우스를 사용하여, 크기와 위치를 조정한다. 자동 ROI 툴을 사용하는 경우 모든 이미지에 대해 동일한 투자 수익 (ROI) 임계 값을 사용합니다. ROI의 크기와 위치를 수동으로 (예를 들어, 원형의 ROI) 정의 된 경우 모든 이미지에 대해 동일한 영역을 사용합니다. </li><li> RO.I. 새 창 팝업을 위로 관찰 측정 &quot;을 클릭합니다. 열을 사용자 정의 (예를 들어, 파일 이름, 동물 번호, 그룹, 실험, 면적, 총 수, 평균 횟수, SD 수, 최소 및 최대 카운트, 면적, 시간 포인트, 프로브 주입 등)의 시간. 사용자 정의 된 설정을 저장합니다. 때 다시ADY 모든 (Ctrl + A)를 선택하고 복사하여 스프레드 시트에 표를 붙여 넣습니다. </li><li> 장소에 ROI를 가진 PNG로 이미지를 내 보냅니다. 저장 한 이미지 파일을 닫습니다. </li></ol></li><li> 정량하는 데 필요한 모든 이미지 파일 -이 절차를 반복합니다 (3.1.7 3.1.6 단계). 스프레드 시트에 모든 ROI의 정량화를 복사합니다. 참고 : 여기에, 결과 등 그룹, 시점, 정렬 할 수 있습니다 통계적으로 분석 하였다. 통계 분석에 대한 ROI를에 생물 발광 신호의 총 수를 사용합니다. </li></ol></li><li> 형광 나노 입자의 근적외선 (NIR) 분석 <ol><li> 소프트웨어의 컨트롤을 사용하여 이미지의 임계치를 조정한다. 참고 :이 양적 결과에 영향을주지 않습니다. </li><li> 시각 신호의 특이성을 평가하기 위해 예 / 엠 720분의 675 및 예 ​​/ 엠 700분의 640에서 촬영 한 이미지를 비교. </li><li> 정량 분석에 대한 예 / 엠 720분의 675에서 촬영 한 화상을 사용하여 (여기 최대 : 680 ㎚). 자동 ROI 도구가 이용 될 수있는 로아를 정의. 자동 ROI 임계 값을 조정하고 모든 이미지를 사용합니다. 로아의 전체 방사 효율을 정량화한다 (단계 3.1 참조). </li></ol></li><li> 단백질 분해 효소에 민감한 프로브의 근적외선 (NIR) 분석 <ol><li> 소프트웨어 제어를 사용하여 이미지의 임계치를 조정한다. 참고 :이 양적 결과에 영향을주지 않습니다. 자동 형광 스펙트럼 unmixing을 수행합니다. unmixing 도구는 이미지 생활에서 구현됩니다. 자동 unmixing은 자동 형광 이미지로 특정하고 720분의 675로 예 / 엠 700분의 640을 사용합니다. </li><li> 혼합되지 않은 이미지를 선택하고 상술 한 바와 같이 ROI를 정의합니다. 정량 및 통계 분석에 대한 ROI를 총 방사 효율을 사용합니다. </li></ol></li></ol>

<ol>
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L., 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=Optical+tomographic+imaging+of+near+infrared+imaging+agents+quantifies+disease+severity+and+immunomodulation+of+experimental+autoimmune+encephalomyelitis+in+vivo.">Optical tomographic imaging of near infrared imaging agents quantifies disease severity and immunomodulation of experimental autoimmune encephalomyelitis in vivo.</a> J Neuroinflammation. 10, (2013).</li><li>Kandagaddala, L. D., Kang, M. J., Chung, B. C., Patterson, T. A., Kwon, O. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+and+activation+of+matrix+metalloproteinase-9+and+NADPH+oxidase+in+tissues+and+plasma+of+experimental+autoimmune+encephalomyelitis+in+mice.">Expression and activation of matrix metalloproteinase-9 and NADPH oxidase in tissues and plasma of experimental autoimmune encephalomyelitis in mice.</a> Exp Toxicol Pathol. 64 (1-2), 109-114 (2012).</li><li>Wang, C., 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=In+situ+fluorescence+imaging+of+myelination.">In situ fluorescence imaging of myelination.</a> J Histochem Cytochem. 58 (7), 611-621 (2010).</li><li>Wang, C., 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=Longitudinal+near-infrared+imaging+of+myelination.">Longitudinal near-infrared imaging of myelination.</a> J Neurosci. 31 (7), 2382-2390 (2011).</li><li>Engelhardt, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+mechanisms+involved+in+T+cell+migration+across+the+blood-brain+barrier.">Molecular mechanisms involved in T cell migration across the blood-brain barrier.</a> J Neural Transm. 113 (4), 477-485 (2006).</li><li>Badawi, A. H., 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=Suppression+of+EAE+and+prevention+of+blood-brain+barrier+breakdown+after+vaccination+with+novel+bifunctional+peptide+inhibitor.">Suppression of EAE and prevention of blood-brain barrier breakdown after vaccination with novel bifunctional peptide inhibitor.</a> Neuropharmacology. 62 (4), 1874-1881 (2012).</li><li>Simmons, S. B., Pierson, E. R., Lee, S. Y., Goverman, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modeling+the+heterogeneity+of+multiple+sclerosis+in+animals.">Modeling the heterogeneity of multiple sclerosis in animals.</a> Trends in immunology. 34 (8), 410-422 (2013).</li><li>Lin, T. H., 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=Diffusion+fMRI+detects+white-matter+dysfunction+in+mice+with+acute+optic+neuritis.">Diffusion fMRI detects white-matter dysfunction in mice with acute optic neuritis.</a> Neurobiol Dis. 67, 1-8 (2014).</li><li>Knier, B., 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=Neutralizing+IL-17+protects+the+optic+nerve+from+autoimmune+pathology+and+prevents+retinal+nerve+fiber+layer+atrophy+during+experimental+autoimmune+encephalomyelitis.">Neutralizing IL-17 protects the optic nerve from autoimmune pathology and prevents retinal nerve fiber layer atrophy during experimental autoimmune encephalomyelitis.</a> J Autoimmun. 56, 34-44 (2015).</li><li>Schellenberg, A. E., Buist, R., Yong, V. W., Del Bigio, M. R., Peeling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Magnetic+resonance+imaging+of+blood-spinal+cord+barrier+disruption+in+mice+with+experimental+autoimmune+encephalomyelitis.">Magnetic resonance imaging of blood-spinal cord barrier disruption in mice with experimental autoimmune encephalomyelitis.</a> Magn Reson Med. 58 (2), 298-305 (2007).</li><li>Mori, Y., 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=Early+pathological+alterations+of+lower+lumbar+cords+detected+by+ultrahigh-field+MRI+in+a+mouse+multiple+sclerosis+model.">Early pathological alterations of lower lumbar cords detected by ultrahigh-field MRI in a mouse multiple sclerosis model.</a> Int Immunol. 26 (2), 93-101 (2014).</li><li>Bittner, 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=Endothelial+TWIK-related+potassium+channel-1+(TREK1)+regulates+immune-cell+trafficking+into+the+CNS.">Endothelial TWIK-related potassium channel-1 (TREK1) regulates immune-cell trafficking into the CNS.</a> Nat Med. 19 (9), 1161-1165 (2013).</li><li>Theien, B. E., 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=Differential+effects+of+treatment+with+a+small-molecule+VLA-4+antagonist+before+and+after+onset+of+relapsing+EAE.">Differential effects of treatment with a small-molecule VLA-4 antagonist before and after onset of relapsing EAE.</a> Blood. 102 (13), 4464-4471 (2003).</li><li>Hawkins, B. T., Davis, T. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+blood-brain+barrier/neurovascular+unit+in+health+and+disease.">The blood-brain barrier/neurovascular unit in health and disease.</a> Pharmacol Rev. 57 (2), 173-185 (2005).</li><li>Coisne, C., Mao, W., Engelhardt, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cutting+edge:+Natalizumab+blocks+adhesion+but+not+initial+contact+of+human+T+cells+to+the+blood-brain+barrier+in+vivo+in+an+animal+model+of+multiple+sclerosis.">Cutting edge: Natalizumab blocks adhesion but not initial contact of human T cells to the blood-brain barrier in vivo in an animal model of multiple sclerosis.</a> J Immunol. 182 (10), 5909-5913 (2009).</li><li>Andresen, V., 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=High-resolution+intravital+microscopy.">High-resolution intravital microscopy.</a> PLoS One. 7 (12), e50915 (2012).</li><li>Bukilica, M., 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=Stress-induced+suppression+of+experimental+allergic+encephalomyelitis+in+the+rat.">Stress-induced suppression of experimental allergic encephalomyelitis in the rat.</a> Int J Neurosci. 59 (1-3), 167-175 (1991).</li></ol>

저자는 더 경쟁 재정적 이해 관계가 없음을 선언합니다.

본 동영상은 SJL / J 마우스의 EAE의 생체 내 이미징의 생물 발광 및 근적외선 형광을위한 기술을 보여줍니다. 우리는 염증에 민감한 프로브를 사용하여 생물 발광 영상은 주로 시신경염를 표시하고 정량화가 EAE 심각도의 임상 평가 및 약물의 효과에 동의 것으로 나타났다. 신호가 척추에 의해 흡수되기 때문에, 생물 발광 촬상 방법 가능성 EAE 현시 (17)의 기본 위치 인 요추 ?...

Multiple sclerosis is caused by the autoimmune-mediated attack and destruction of the myelin sheath in the brain and the spinal cord1. With an overall incidence of about 3.6 cases per 100,000 people a year in women and about 2.0 in men, MS is the second most common cause of neurological disability in young adults, after traumatic injuries2,3. The disease pathology is contributed to by genetic and environmental factors4 but is still not completely understood. Autoreactive T lymphocytes enter the central nervous system and trigger an inflammatory cascade that causes focal infiltrates in the white matter of the brain, spinal cord, and optic nerve. In most cases, these infiltrates are initially reversible, but persistence increases with the number of relapses. A number of rodent models have been developed to study the pathology of the disease. The relapsing-remitting EAE in SJL/J mice and the primary-progressive EAE in C57BL6 mice are the most popular models.
The clinical EAE scores, which describe the extent of the motor function deficits, and body weight are the gold standards to assess EAE severity. These clinical signs agree with the extent of immune cell infiltration and myelin destruction in the spinal cord and moderately predict drug treatment efficacy in humans5. However, these signs mainly reflect the destruction of the ventral fiber tracts in the spinal cord. Presently, there is no easy, non-invasive, reliable, and reproducible method to assess in vivo brain infiltration and optic neuritis in living mice. 
The in vivo imaging agrees with the 3 "R" principles of Russel and Burch (1959), which claim a Replacement, Reduction, and Refinement of animal experiments6, because imaging increases the readouts of one animal at several time points and allows for a reduction of the overall numbers. Presently, inflammation or myelin status is mainly assessed ex vivo via immunohistochemistry, FACS-analysis, or different molecular biological methods7, all requiring euthanized mice at specific time points. 
A number of in vivo imaging system probes have been developed to assess inflammation in the skin, joints, and vascular system. The techniques rely on the activation of bioluminescent or near-infrared fluorescent substrates by tissue peroxidases, including myeloperoxidase (MPO), matrix metalloproteinases (MMPs)8, and cathepsins9 or cyclooxygenase2. These probes have been mainly validated in models of arthritis or atherosclerosis9,10. A cathepsin-sensitive probe has also been used for fluorescence molecular tomographic imaging of EAE11. MMPs, particularly MMP2 and MMP9, contribute to the protease-mediated BBB disruption in EAE and are upregulated at sites of immune cell infiltration12, suggesting that these probes may be useful for EAE imaging. The same holds true for peroxidase or cathepsin-based probes. Technically, imaging of inflammation in the brain or spinal cord is substantially more challenging because the skull or spine absorb bioluminescent and near-infrared signals. 
In addition to inflammation indicators, fluorescent chemicals have been described, which specifically bind to myelin and may allow for quantification of myelination13. A near-infrared fluorescent probe, 3,3'-diethylthiatricarbocyanine iodide (DBT), was found to specifically bind to myelinated fibers and was validated as a quantitative tool in mouse models of primary myelination defects and in cuprizone-evoked demyelination14. In EAE, the DBT signal was rather increased, reflecting the inflammation of the myelin fibers5. 
An additional hallmark of EAE and MS is the BBB breakdown, resulting in increased vascular permeability and the extravasation of blood cells, extracellular fluid, and macromolecules into the CNS parenchyma. This can lead to edema, inflammation, oligodendrocyte damage, and, eventually, demyelination15,16. Hence, visualization of the BBB leak using fluorescent probes, such as fluorochrome-labeled bovine serum albumin5, which normally distribute very slowly from blood to tissue, may be useful to assess EAE.
In the present study, we have assessed the usefulness of different probes in EAE and show the procedure for the most reliable and robust bioluminescent technique. In addition, we discuss the pros and cons of near-infrared probes for MMP activity and BBB integrity.

<table border="0" cellpadding="0" cellspacing="0" width="1287">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">AngioSpark-680</td>
			<td style="width:267px;">Perkin Elmer, Inc., Waltham, USA</td>
			<td style="width:119px;">NEV10149</td>
			<td style="width:636px;">Imaging probe, pegylated nanoparticles, useful for imaging of blood brain barrier integrity</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">MMP-sense 680</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td>NEV10126</td>
			<td>Imaging probe, activatable by matrix metalloproteinases, useful for imaging of inflammation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">XenoLight RediJect Inflammation Probe</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td>760535</td>
			<td>Imaging probe, activatable by oxidases, useful for imaging of inflammation</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">PLP139-151/CFA emulsion&#160;</td>
			<td>Hooke Labs, St Lawrence, MA</td>
			<td>EK-0123</td>
			<td>EAE induction kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Pertussis Toxin</td>
			<td>Hooke Labs, St Lawrence, MA</td>
			<td>EK-0123</td>
			<td>EAE induction kit</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">IVIS Lumina Spectrum</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td style="width:119px;"></td>
			<td>Bioluminescence and Infrared Imaging System</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">LivingImage 4.5 software&#160;</td>
			<td>Perkin Elmer, Inc., Waltham, USA</td>
			<td>CLS136334</td>
			<td>IVIS analysis software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:265px;">Isoflurane</td>
			<td style="width:267px;">Abbott Labs, Illinois, USA</td>
			<td>26675-46-7</td>
			<td>Anaesthetic</td>
		</tr>
	</tbody>
</table>

bioluminescence and near-infrared imaging of optic neuritis and brain inflammation in the eae model of multiple sclerosis in mice

SJL / J 쥐에 실험자가 면역 뇌척수염 (EAE)는 재발 - 완화 성 다발성 경화증 (RRMS)의 모델이다. 운동 기능 적자를 설명하는 임상 EAE 점수는 척수의 면역 매개 염증의 기본 판독입니다. 그러나 점수와 체중은 뇌의 염증과 시신경염의 생체 내 평가를 허용하지 않습니다. 후자는 2/3 MS 환자에서 초기 자주 표현된다. 여기서는 생체 촬상 시스템을 이용하여 살아있는 쥐 생물 발광과 EAE는 시신경염을 유발 평가 근적외선 라이브 영상, 뇌 염증 및 혈액 - 뇌 장벽 (BBB) 중단하는 방법을 보여준다. 산화 효소에 의해 활성화 생물 발광 기판은 주로 시신경염 나타났다. 신호는 특정이었고, 임상 점수를 병렬 약물 효과 및 질병 시간 과정의 시각화를 허용했다. vasculatur 내에 남아 페 길화 된 형광 나노 입자장시간 E는 BBB의 무결성을 평가하기 위해 사용되었다. 근적외선 영상은 질병의 피크에서 BBB 누수를 한 것으로 밝혀졌습니다. 신호는 눈 주위 강했다. 매트릭스 메탈 용 근적외선 기판 EAE - 유발 된 염증을 평가하기 위해 사용되었다. 자동 형광 정량 unmixing 스펙트럼을 요구 신호 간섭. 전반적으로, 생물 발광 이미징 EAE 관련 시신경염 약물 효과를 평가하기위한 신뢰성있는 방법이고 신호 특이성 견고성 정량의 용이성 및 비용면에서 근적외선 기술보다 우수 하였다.

시신경염의 생물 발광의 시간 코스 염증 프로브의 생물 발광 신호는 눈 주위 강한이었고 시신경염와 EAE 마우스에서만 일어났다. 신호는 비 EAE 마우스 나 염증 프로브를 주입하지 생쥐도 발생 하였다. 쥐를 복구 할 때 신호가 사라졌다. 따라서, 신호 시신경염 특정하고, 상기 신호의 피크 임상 EAE 스코어의 피크 평...

Watch this Scientific Journal Video about Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice at JoVE.com

Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

우리는 생체 살아있는 생물 발광 및 근적외선 SJL / J 마우스에서 다발성 경화증에 대한 실험자가 면역 뇌척수염 (EAE) 모델에서 시신경염과 뇌염의 이미징 기술을 보여준다.

마우스의 다발성 경화증의 EAE 모델에서 시신경염 및 뇌 염증의 생물 발광 및 근적외선 영상

Experimental autoimmune encephalomyelitis (EAE) in SJL/J mice is a model for relapsing-remitting multiple sclerosis (RRMS). Clinical EAE scores describing ...

bioluminescence-near-infrared-imaging-optic-neuritis-brain

Research

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Medicine

9.1K 조회수.  University Hospital Frankfurt.  우리는 생체 살아있는 생물 발광 및 근적외선 SJL / J 마우스에서 다발성 경화증에 대한 실험자가 면역 뇌척수염 (EAE) 모델에서 시신경염과 뇌염의 이미징 기술을 보여준다. 

비디오: Bioluminescence and Near-infrared Imaging of Optic Neuritis and Brain Inflammation in the EAE Model of Multiple Sclerosis in Mice

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading to an accumulation of lipid-laden intra-luminal plaque, smooth muscle cell proliferation and progressive narrowing of the vessel lumen. The formation of such vulnerable plaques prone to rupture underlies the majority of cases of acute myocardial infarction. The complex molecular and cellular inflammatory cascade is orchestrated by the recruitment of T lymphocytes and macrophages and their paracrine effects on endothelial and smooth muscle cells.1
Molecular imaging in atherosclerosis has evolved into an important clinical and research tool that allows in vivo visualization of inflammation and other biological processes. Several recent examples demonstrate the ability to detect high-risk plaques in patients, and assess the effects of pharmacotherapeutics in atherosclerosis.4 While a number of molecular imaging approaches (in particular MRI and PET) can image biological aspects of large vessels such as the carotid arteries, scant options exist for imaging of coronary arteries.2 The advent of high-resolution optical imaging strategies, in particular near-infrared fluorescence (NIRF), coupled with activatable fluorescent probes, have enhanced sensitivity and led to the development of new intravascular strategies to improve biological imaging of human coronary atherosclerosis.
Near infrared fluorescence (NIRF) molecular imaging utilizes excitation light with a defined band width (650-900 nm) as a source of photons that, when delivered to an optical contrast agent or fluorescent probe, emits fluorescence in the NIR window that can be detected using an appropriate emission filter and a high sensitivity charge-coupled camera. As opposed to visible light, NIR light penetrates deeply into tissue, is markedly less attenuated by endogenous photon absorbers such as hemoglobin, lipid and water, and enables high target-to-background ratios due to reduced autofluorescence in the NIR window. Imaging within the NIR 'window' can substantially improve the potential for in vivo imaging.2,5
Inflammatory cysteine proteases have been well studied using activatable NIRF probes10, and play important roles in atherogenesis. Via degradation of the extracellular matrix, cysteine proteases contribute importantly to the progression and complications of atherosclerosis8. In particular, the cysteine protease, cathepsin B, is highly expressed and colocalizes with macrophages in experimental murine, rabbit, and human atheromata.3,6,7 In addition, cathepsin B activity in plaques can be sensed in vivo utilizing a previously described 1-D intravascular near-infrared fluorescence technology6, in conjunction with an injectable nanosensor agent that consists of a poly-lysine polymer backbone derivatized with multiple NIR fluorochromes (VM110/Prosense750, ex/em 750/780nm, VisEn Medical, Woburn, MA) that results in strong intramolecular quenching at baseline.10 Following targeted enzymatic cleavage by cysteine proteases such as cathepsin B (known to colocalize with plaque macrophages), the fluorochromes separate, resulting in substantial amplification of the NIRF signal. Intravascular detection of NIR fluorescence signal by the utilized novel 2D intravascular NIRF catheter now enables high-resolution, geometrically accurate in vivo detection of cathepsin B activity in inflamed plaque.
In vivo molecular imaging of atherosclerosis using catheter-based 2D NIRF imaging, as opposed to a prior 1-D spectroscopic approach,6 is a novel and promising tool that utilizes augmented protease activity in macrophage-rich plaque to detect vascular inflammation.11,12 The following research protocol describes the use of an intravascular 2-dimensional NIRF catheter to image and characterize plaque structure utilizing key aspects of plaque biology. It is a translatable platform that when integrated with existing clinical imaging technologies including angiography and intravascular ultrasound (IVUS), offers a unique and novel integrated multimodal molecular imaging technique that distinguishes inflammatory atheromata, and allows detection of intravascular NIRF signals in human-sized coronary arteries.

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

We detail a new near-infrared fluorescence (NIRF) catheter for 2-dimensional intravascular molecular imaging of plaque biology in vivo. The NIRF catheter can visualize key biological processes such as inflammation by reporting on the presence of plaque-avid activatable and targeted NIR fluorochromes. The catheter utilizes clinical engineering and power requirements and is targeted for application in human coronary arteries. The following research study describes a multimodal imaging strategy that utilizes a novel in vivo intravascular NIRF catheter to image and quantify inflammatory plaque in proteolytically active inflamed rabbit atheromata.

염증성 플라크의 적외선 형광 (NIRF) Intravascular 분자 이미징, Multimodal 접근 가까운 생체내에서 죽상 경화증의 영상을

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

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In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There is little knowledge about in situ, in vivo, tumor hypoxia during intracranial development of malignant brain tumors because of lack of efficient means to monitor it in these deep-seated orthotopic tumors. Bioluminescence imaging (BLI), based on the detection of light emitted by living cells expressing a luciferase gene, has been rapidly adopted for cancer research, in particular, to evaluate tumor growth or tumor size changes in response to treatment in preclinical animal studies. Moreover, by expressing a reporter gene under the control of a promoter sequence, the specific gene expression can be monitored non-invasively by BLI. Under hypoxic stress, signaling responses are mediated mainly via the hypoxia inducible factor-1&alpha; (HIF-1&alpha;) to drive transcription of various genes. Therefore, we have used a HIF-1&alpha; reporter construct, 5HRE-ODD-luc, stably transfected into human breast cancer MDA-MB231 cells (MDA-MB231/5HRE-ODD-luc). In vitro HIF-1&alpha; bioluminescence assay is performed by incubating the transfected cells in a hypoxic chamber (0.1% O2) for 24 hr before BLI, while the cells in normoxia (21% O2) serve as a control. Significantly higher photon flux observed for the cells under hypoxia suggests an increased HIF-1&alpha; binding to its promoter (HRE elements), as compared to those in normoxia. Cells are injected directly into the mouse brain to establish a breast cancer brain metastasis model. In vivo bioluminescence imaging of tumor hypoxia dynamics is initiated 2 wks after implantation and repeated once a week. BLI reveals increasing light signals from the brain as the tumor progresses, indicating increased intracranial tumor hypoxia. Histological and immunohistochemical studies are used to confirm the in vivo imaging results. Here, we will introduce approaches of in vitro HIF-1&alpha; bioluminescence assay, surgical establishment of a breast cancer brain metastasis in a nude mouse and application of in vivo bioluminescence imaging to monitor intracranial tumor hypoxia.

In vivo Bioluminescence Imaging of Tumor Hypoxia Dynamics of Breast Cancer Brain Metastasis in a Mouse Model

Bioluminescence imaging of hypoxia inducible factor-1&alpha; activity is applied to monitor intracranial tumor hypoxia development in a breast cancer brain metastasis mouse model.

유방암 뇌 전이의 종양 Hypoxia 역학 생체내의 Bioluminescence 이미징에서는 마우스 모델에서

It is well recognized that tumor hypoxia plays an important role in promoting malignant progression and affecting therapeutic response negatively. There ...

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis (ALS)

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 years for the sporadic disease and around 50 years for the familial disease. Due to its progressive course, 50% of the patients die within 30 months of symptom onset. In order to evaluate novel treatment options for this disease, genetic mouse models of ALS have been generated based on human familial mutations in the SOD gene, such as the SOD1 (G93A) mutation. Most important aspects that have to be evaluated in the model are overall survival, clinical course and motor function. Here, we demonstrate the clinical evaluation, show the conduction of two behavioural motor tests and provide quantitative scoring systems for all parameters. Because an in depth analysis of the ALS mouse model usually requires an immunohistochemical examination of the spinal cord, we demonstrate its preparation in detail applying the dorsal laminectomy method. Exemplary histological findings are demonstrated. The comprehensive application of the depicted examination methods in studies on the mouse model of ALS will enable the researcher to reliably test future therapeutic options which can provide a basis for later human clinical trials.

Clinical Testing and Spinal Cord Removal in a Mouse Model for Amyotrophic Lateral Sclerosis ALS

A mouse model for amyotrophic lateral sclerosis (ALS) is examined clinically and behaviorally. As a prerequisite for an accompanying immunohistological analysis the preparation of the spinal cord is depicted in detail.

루게릭병에 대한 마우스 모델 (ALS)의 임상 시험 및 척수 제거

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder resulting in progressive degeneration of motoneurons. Peak of onset is around 60 ...

Dynamic Visual Tests to Identify and Quantify Visual Damage and Repair Following Demyelination in Optic Neuritis Patients

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to assess changes in myelination at the central nervous system (CNS). However, standard measurements, including routine visual tests and MRI scans, are not sensitive enough for this purpose. We present two visual tests addressing dynamic monocular and binocular functions which may closely associate with the extent of myelination along visual pathways. These include Object From Motion (OFM) extraction and Time-constrained stereo protocols. In the OFM test, an array of dots compose an object, by moving the dots within the image rightward while moving the dots outside the image leftward or vice versa. The dot pattern generates a camouflaged object that cannot be detected when the dots are stationary or moving as a whole. Importantly, object recognition is critically dependent on motion perception. In the Time-constrained Stereo protocol, spatially disparate images are presented for a limited length of time, challenging binocular 3-dimensional integration in time. Both tests are appropriate for clinical usage and provide a simple, yet powerful, way to identify and quantify processes of demyelination and remyelination along visual pathways. These protocols may be efficient to diagnose and follow optic neuritis and multiple sclerosis patients.
In the diagnostic process, these protocols may reveal visual deficits that cannot be identified via current standard visual measurements. Moreover, these protocols sensitively identify the basis of the currently unexplained continued visual complaints of patients following recovery of visual acuity. In the longitudinal follow up course, the protocols can be used as a sensitive marker of demyelinating and remyelinating processes along time. These protocols may therefore be used to evaluate the efficacy of current and evolving therapeutic strategies, targeting myelination of the CNS.

Object From Motion (OFM) and Time-constrained stereo protocols are sensitive tools to identify monocular and binocular dynamic visual function deficits, which are uniquely affected in optic neuritis patients. Furthermore, these tests may be used as quantitative noninvasive tools to assess the extent of myelination along visual pathways.

시신경염 환자에서 탈수 초화 다음 Visual 손상 및 수리를 확인하고 정량화하는 동적 인 비주얼 테스트

In order to follow optic neuritis patients and evaluate the effectiveness of their treatment, a handy, accurate and quantifiable tool is required to ...

The Multiple Sclerosis Performance Test (MSPT): An iPad-Based Disability Assessment Tool

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report a new test, the Multiple Sclerosis Performance Test (MSPT), which represents a new approach to quantifying MS related disability. The MSPT takes advantage of advances in computer technology, information technology, biomechanics, and clinical measurement science. The resulting MSPT represents a computer-based platform for precise, valid measurement of MS severity. Based on, but extending the Multiple Sclerosis Functional Composite (MSFC), the MSPT provides precise, quantitative data on walking speed, balance, manual dexterity, visual function, and cognitive processing speed. The MSPT was tested by 51 MS patients and 49 healthy controls (HC). MSPT scores were highly reproducible, correlated strongly with technician-administered test scores, discriminated MS from HC and severe from mild MS, and correlated with patient reported outcomes. Measures of reliability, sensitivity, and clinical meaning for MSPT scores were favorable compared with technician-based testing. The MSPT is a potentially transformative approach for collecting MS disability outcome data for patient care and research. Because the testing is computer-based, test performance can be analyzed in traditional or novel ways and data can be directly entered into research or clinical databases. The MSPT could be widely disseminated to clinicians in practice settings who are not connected to clinical trial performance sites or who are practicing in rural settings, drastically improving access to clinical trials for clinicians and patients. The MSPT could be adapted to out of clinic settings, like the patient&#8217;s home, thereby providing more meaningful real world data. The MSPT represents a new paradigm for neuroperformance testing. This method could have the same transformative effect on clinical care and research in MS as standardized computer-adapted testing has had in the education field, with clear potential to accelerate progress in clinical care and research.

The Multiple Sclerosis Performance Test MSPT: An iPad-Based Disability Assessment Tool

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report methodologic details on a new test, the Multiple Sclerosis Performance Test (MSPT). This new approach to the objective of quantification of MS related disability provides a computer-based platform for precise, valid measurement of MS severity.

다발성 경화증 성능 테스트 (MSPT) : 아이 패드 기반 장애 평가 도구

Precise measurement of neurological and neuropsychological impairment and disability in multiple sclerosis is challenging. We report a new test, the ...

Th17 Inflammation Model of Oropharyngeal Candidiasis in Immunodeficient Mice

Oropharyngeal Candidiasis (OPC) disease is caused not only due to the lack of host immune resistance, but also the absence of appropriate regulation of infection-induced immunopathology. Although Th17 cells are implicated in antifungal defense, their role in immunopathology is unclear. This study presents a method for establishing oral Th17 immunopathology associated with oral candidal infection in immunodeficient mice. The method is based on reconstituting lymphopenic mice with in vitro cultured Th17 cells, followed by oral infection with Candida albicans (C. albicans). Results show that unrestrained Th17 cells result in inflammation and pathology, and is associated with several measurable read-outs including weight loss, pro-inflammatory cytokine production, tongue histopathology and mortality, showing that this model may be valuable in studying OPC immunopathology. Adoptive transfer of regulatory cells (Tregs) controls and reduces the inflammatory response, showing that this model can be used to test new strategies to counteract oral inflammation. This model may also be applicable in studying oral Th17 immunopathology in general in the context of other oral diseases.

Although Candida infection models are available to study host immune resistance, a model to study T cell mediated immunopathology in the context of Candida infection is absent. Here we describe a method to establish Th17 immunopathology associated with oral Candida infection in immunodeficient mice.

면역 결핍 마우스의 구인두 칸디다증의 Th17 염증 모델

Oropharyngeal Candidiasis (OPC) disease is caused not only due to the lack of host immune resistance, but also the absence of appropriate regulation of ...

A Method of Trigonometric Modelling of Seasonal Variation Demonstrated with Multiple Sclerosis Relapse Data

This report describes a novel Stata-based application of trigonometric regression modelling to 55 years of multiple sclerosis relapse data from 46 clinical centers across 20 countries located in both hemispheres. Central to the success of this method was the strategic use of plot analysis to guide and corroborate the statistical regression modelling. Initial plot analysis was necessary for establishing realistic hypotheses regarding the presence and structural form of seasonal and latitudinal influences on relapse probability and then testing the performance of the resultant models. Trigonometric regression was then necessary to quantify these relationships, adjust for important confounders and provide a measure of certainty as to how plausible these associations were. Synchronization of graphing techniques with regression modelling permitted a systematic refinement of models until best-fit convergence was achieved, enabling novel inferences to be made regarding the independent influence of both season and latitude in predicting relapse onset timing in MS. These methods have the potential for application across other complex disease and epidemiological phenomena suspected or known to vary systematically with season and/or geographic location.

Combining plot analysis with trigonometric regression is a robust method for exploring complex, cyclical phenomena such as relapse onset timing in multiple sclerosis (MS). This method enabled unbiased characterisation of seasonal trends in relapse onset permitting novel inferences around the influence of seasonal variation, ultraviolet radiation (UVR) and latitude.

다발성 경화증 재발 데이터로 입증 된 계절 변화의 삼각 모델링하는 방법

This report describes a novel Stata-based application of trigonometric regression modelling to  55 years of multiple sclerosis relapse data from 46 ...

A Protocol for the Use of Remotely-Supervised Transcranial Direct Current Stimulation (tDCS) in Multiple Sclerosis (MS)

Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique that uses low amplitude direct currents to alter cortical excitability. With well-established safety and tolerability, tDCS has been found to have the potential to ameliorate symptoms such as depression and pain in a range of conditions as well as to enhance outcomes of cognitive and physical training. However, effects are cumulative, requiring treatments that can span weeks or months and frequent, repeated visits to the clinic. The cost in terms of time and travel is often prohibitive for many participants, and ultimately limits real-world access. 
Following guidelines for remote tDCS application, we propose a protocol that would allow remote (in-home) participation that uses specially-designed devices for supervised use with materials modified for patient use, and real-time monitoring through a telemedicine video conferencing platform. We have developed structured training procedures and clear, detailed instructional materials to allow for self- or proxy-administration while supervised remotely in real-time. The protocol is designed to have a series of checkpoints, addressing attendance and tolerability of the session, to be met in order to continue to the next step. The feasibility of this protocol was then piloted for clinical use in an open label study of remotely-supervised tDCS in multiple sclerosis (MS). This protocol can be widely used for clinical study of tDCS.

A Protocol for the Use of Remotely-Supervised Transcranial Direct Current Stimulation tDCS in Multiple Sclerosis MS

The goal of this pilot study is to describe a protocol for the remotely-supervised delivery of transcranial direct current stimulation (tDCS) so that the procedure maintains standards of in-clinic practice, including safety, reproducibility, and tolerability. The feasibility of this protocol was tested in participants with multiple sclerosis (MS).

다발성 경화증에 원격으로 감독 경 두개 직류 자극 (tDCS)의 사용을위한 프로토콜 (MS)

Transcranial direct current stimulation (tDCS) is a noninvasive brain stimulation technique that uses low amplitude direct currents to alter cortical ...

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of stroke than relying on indirect measurements, and will be a potent and robust vascular research tool. We use an optical imaging approach that labels thrombi with a molecular imaging thrombus marker&#160;&#8212; a Cy5.5 near-infrared fluorescent (NIRF) probe that is covalently linked to the fibrin strands of the thrombus by the fibrin-crosslinking enzymatic action of activated coagulation factor XIIIa during the process of clot maturation. A micro-computed tomography (microCT)-based approach uses thrombus-seeking gold nanoparticles (AuNPs) functionalized to target the major component of the clot: fibrin. This paper describes a detailed protocol for the combined in vivo microCT and ex vivo NIRF imaging of thromboemboli in a mouse model of embolic stroke. We show that in vivo microCT and fibrin-targeted glycol-chitosan AuNPs (fib-GC-AuNPs) can be used for visualizing both in situ thrombi and cerebral embolic thrombi. We also describe the use of in vivo microCT-based direct thrombus imaging to serially monitor the therapeutic effects of tissue plasminogen activator-mediated thrombolysis. After the last imaging session, we demonstrate by ex vivo NIRF imaging the extent and the distribution of residual thromboemboli in the brain. Finally, we describe quantitative image analyses of microCT and NIRF imaging data. The combined technique of direct thrombus imaging allows two independent methods of thrombus visualization to be compared: the area of thrombus-related fluorescent signal on ex vivo NIRF imaging vs. the volume of hyperdense microCT thrombi in vivo.

This protocol describes the application of combined near-infrared fluorescent (NIRF) imaging and micro-computed tomography (microCT) for visualizing cerebral thromboemboli. This technique allows the quantification of thrombus burden and evolution. The NIRF imaging technique visualizes fluorescently labeled thrombus in excised brain, while the microCT technique visualizes thrombus inside living animals using gold-nanoparticles.

직접 대뇌 Thromboemboli 시각화를위한 결합 된 근적외선 형광 이미징 및 마이크로 계산 된 단층 촬영

Direct thrombus imaging visualizes the root cause of thromboembolic infarction. Being able to image thrombus directly allows far better investigation of ...

Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic demand. Reactive hyperemia (in response to a brief period of tissue ischemia) is an independent predictor of cardiovascular events and provides important insight into vascular health and vasodilatory capacity. Skeletal muscle oxidative capacity is equally important in health and disease, as it determines the energy supply for myocellular processes. Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess each of these major clinical endpoints (reactive hyperemia, neurovascular coupling, and muscle oxidative capacity) during a single clinic or laboratory visit. Unlike Doppler ultrasound, magnetic resonance images/spectroscopy, or invasive catheter-based flow measurements or muscle biopsies, our approach is less operator-dependent, low-cost, and completely non-invasive. Representative data from our lab taken together with summary data from previously published literature illustrate the utility of each of these end-points. Once this technique is mastered, application to clinical populations will provide important mechanistic insight into exercise intolerance and cardiovascular dysfunction.

Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia, neurovascular coupling and skeletal muscle oxidative capacity in a single clinic or laboratory visit.

골격 근육 혈관 결합, 산화 용량 및 Microvascular 함수 '원스톱 쇼핑' 근처-적외선 분광학

Exercise represents a major hemodynamic stress that demands a highly coordinated neurovascular response in order to match oxygen delivery to metabolic ...