Bu çalışma, Ulusal Kalp, Akciğer ve Kan Enstitüsü Hibe Ödülü Numaraları R01HL096593 ve K23HL72825 tarafından desteklenmiştir. Içeriği yalnızca Thomas Jefferson Üniversitesi sorumluluğundadır ve mutlaka Ulusal Kalp, Akciğer ve Kan Enstitüsü ve Ulusal Sağlık Enstitüleri resmi görüşlerini temsil etmemektedir.

Olgu Sunumu (gerekli ise): NA, bu hala deneysel bir araştırma prosedürü, henüz klinik olarak kullanılmamaktadır. Tanı, Değerlendirme ve Planı (gerekli, varsa): NA, bu hala deneysel bir araştırma prosedürü, henüz klinik olarak kullanılmamaktadır. Prosedür (gerekli): Bu bölüm aşağıdaki kurallara uygun, ilgili prosedürün adım adım açıklama içermelidir. 1. Kapiler Mikroskopi (Şekil 1) <ol><li> Bizim kapillaroskopisi tekniği Serne ve arkadaşları 1 uyarlanmıştır. Kollajen vasküler hastalığı bilinen kapiller değişiklikler 2 üretir çünkü bu işlem için bir dışlama ölçütü, kollajen vasküler hastalıktır. </li><li> En az 10 saatlik gece açlığının ve oturarak dinlenme 20 dakika sonra, mikrovasküler ölçümler 7 arasında bir yarım saat için yapılmaktadırve 11 kalp seviyesinde oturma pozisyonunda ve sol elinde konu ile, sessiz, ısı kontrollü odası (21,5-22,5 ° C arasında muhafaza) içinde, duyuyorum. </li><li> Üçüncü parmak dorsal deri tırnak dibi kılcal (Olympus, Center Valley, Pensilvanya) stereomikroskopta görselleştirildiği, dijital kamera (Model numarası IN-1400: Tanı Cihazları; Sterling Heights, MI) monokrom 4 megapiksel SPOT Insight ile bağlantılı, ve bir dizüstü bilgisayar (Dell Latitude D600: Dell, Austin, Teksas). Hareketi sınırlamak için, sol el ve önkol gevşek katlanmış bir battaniye ile kaplı, ve mikroskop dibinde yerleştirilmiş başka katlanmış battaniye üzerinde dinlenmiş. </li><li> Tırnak yatağı aydınlatma 250 W halojen fiber optik lamba (KL 2500LCD:; Elmsford, NY Schott-Fostec); ile elde edilir tamamlayıcı 150W fiber optik halojen ışık kaynağı (B &amp; B Mikroskoplar, Ltd, Warrendale, Pensilvanya) ek aydınlatma kullanılır koyu pigmentli bireylerde. </li><li> Kılcal, görselleştirmek için3.2x objektif (Olympus 3.2/0.07) 38.4x toplam sistem büyütme kullanılır. </li><li> Kamerayla birlikte verilen SPOT görüntüleme yazılımı, kılcal fotoğraflarda açık / koyu kontrast kullanma bütün deneklerde kılcal damarların görünürlüğünü maksimize etmek için aynı standart SPOT yazılım işlevi (aydınlık ve karanlık seviyesinin germe) kullanılarak geliştirilmiştir. </li><li> Kapiller yoğunluk ölçmek için, dijital fotomikrografı postocclusive reaktif hiperemi sırasında, dinlenme başlangıçta, üç aşamadan her sırasında her 3-5 sn alınır ve venöz oklüzyon sırasında uygulanır. (A) dinlenme Başlangıçta, fotomikrografı kılcal istirahatte perfüze algılamak için üç dakikalık bir süre içinde alınır. Aşağıdaki gibidir (b) postocclusive reaktif hiperemi sırasında fotomikrografik işlevsel perfüze kapiller (taban artı rezerv kılcal damarlar) ölçmek için alınır. Önce sol üst kol üzerinde bir oklüzyon manşet 10 dakika sistolik basıncının üzerinde 40 mm Hg şişirilir. Fotomikrografı sonra inci sırasında alınıre ilk dakikada hemen tüm işlevsel perfüze kapiller görselleştirme, arteriyel oklüzyon yayınlanması sonrasında. Reaktif hiperemi sonrası Alt kapiller yoğunluk engelli fonksiyonel kapiller tutulduğunu gösterir ve bu nedenle fonksiyonel seyrelme. (C) venöz oklüzyon fotomikrografı sırasında hem perfüze (aktif kırmızı kan hücresi (RBC) hareketi ile) ve aşağıdaki gibi kılcal 3 (durgun, hareketsiz eritrositler ile dolu) nonperfused içerir maksimal kapiller yoğunluk, ölçmek için alınır. Postocclusive reaktif hiperemi işlemden sonra kalan on dakika sonra, kolluğu pasif tüm patent kılcal bugünkü içine kan zorlayarak ve fotomikrografları bu süre içinde alınmış, 60 sn için 60 mm Hg şişirilir. Maksimal kapiller yoğunluk içerdiğinden yapısal mevcut tüm kılcal, maksimal kapiller yoğunluğunda azalma yapısal seyrelme gösterir. </li><li> Kapiler yoğunluğu kare binde kılcal sayısı olarak tanımlanırtırnak dibi cilt limeter ve en hareketi tarafından çarpıtılmıştır dört en net odaklı görüntüler elde dört ölçümün ortalaması olarak hesaplanır. Bizim çalışmalarımızda, kılcal sayımları için tipik değerleri (kılcal / mm 2) venöz oklüzyon için post-iskemik ve 90-105 için temel, 65-90 için 55-80 olmuştur. Yüzde kılcal işe değerleri% 5 ve% 25 (~% 10-15 ortalama) arasında ve% 70 ve normotansiflerden daha hipertansiflerde daha düşük olan değerler% 95 (~% 80-90 ortalama), arasındaki kılcal perfüze yüzde genellikle vardır. Sayma işleminin tekrarlanabilirliği 10 farklı konuların fotoğraflarını bağımsız manuel değerlendirmeler (Şekil 2) gerçekleştirilen üç gözlemci ile doğrulandı. Gözlemciler bu konuların kimlik ve kan basıncı bilmeyen. Eğitim sonrasında bağımsız olarak gerçekleştirilir sonraki sayıları anlaşması yüksek düzeyde gösterdi. Ortalama hakemlerarası ve intra-arası farklılıkların orde vardı2-3 kılcal / mm 2 r ve sınıf içi korelasyon katsayıları 0.90 daha tüm fazlaydı. Kapillaroskopisi önlemlerin kısa vadeli varyasyonun hakemlerarası ve intra-arası farklılıklar (yaklaşık 2 kılcal / mm 2) olarak büyüklükte aynı sırada olduğunu, ancak 2-3 yılda gözlenen uzun vadeli varyasyonun büyük büyüklükte bir sipariş oldu (yaklaşık 15 kılcal / mm 2 &#39;), uzunlamasına değişiklikler kolayca değerlendirici varyasyon ayırt edilebileceğini göstermektedir. İki kapiller fonksiyon ölçümlerini Güvenilirlik (sınıf içi korelasyon yüzde kılcal alımı için katsayısı = 0.84 ve yüzde perfüze kapiller için 0.82) de yüksek oldu. </li><li> Araştırmacılar şimdi Image-Pro Plus görüntüleme yazılımı (Şekil 3: Sürüm 6.2, Medya Sibernetik, Inc, Bethesda, MD) kullanılarak kapiller yoğunluk hesaplamak için bir bilgisayar tabanlı yöntemi kullanmaktadır. Bazal, post-iskemik ve veno arasındaki Pearson korelasyon10 konularda yazılım ve ilgili kılavuzu sayımları ile yapılan bize tıkanıklığı sayıları iki yöntem arasında makul bir anlaşma gösteren, (tümü için p &lt;0.05) sırasıyla 0.78, 0.78 ve 0.71 idi. Bilgisayar tabanlı sayıları Güvenilirlik kılavuzu sayıları biraz daha düşüktür, ancak (venöz oklüzyon için başlangıç ​​için sınıf içi korelasyon katsayısı = 0.91, post-iskemik için 0.86 ve 0.84) hala yüksektir. Biz de şu anda yayına hazırlanıyor hipertansiyon dahil olmak üzere birden fazla kardiyovasküler risk faktörleri ile otomatik sayım ilişki gösteren yayınlanmamış veri var. </li><li> Tablo 1 kapiller yoğunluk ölçümleri ve hesaplamaları özetlemektedir. Yüzde kılcal işe maksimal kapiller yoğunluk tarafından postocclusive reaktif hiperemi (postocclusive reaktif hiperemi kapiller yoğunluk eksi başlangıçta kapiller yoğunluk) ile oluşturulan kapiller yoğunluğunda artış, (pasif veno sırasında gözlenen bölünmesiyle değerlendirilirBizi oklüzyonu). Yüzde kılcal (işlevsel aktif) perfüze tüm kılcal mevcut oranını temsil ve maksimal kapiller yoğunluğu postocclusive reaktif hiperemi kapiller yoğunluk bölünmesiyle hesaplanır perfüze. Yüzde kılcal işe alım ve yüzde Hem kılcal fonksiyonel kılcal sayısını yansıtmaktadır perfüze. Bu önlemler için düşük değerlerin fonksiyonel kapiller seyrelme göstermektedir. </li></ol> 2. Endotel Fonksiyonu Değerlendirme <ol><li> Endotel fonksiyonu vazodilatasyon indüklemek için reaktif hiperemi endotele bağımlı uyarıcı kullanan Sivertsson, 4 yöntemine göre, önkol kan akışının invaziv olmayan pletismografi ölçümleri kullanılarak, reaktif hiperemi postocclusive önce ve sonra ölçülür. </li><li> Onun istirahat süresini Mağrası döngü gerçekleştirilir ötesinde sırtüstü kalan 10 dk sonrasında oturma pozisyonunda konu ile, bir cıva kauçuk strain gauge 10% gerginantekübital fossa altına d öznenin önkol 5 cm. </li><li> Sırayla bir kayıt cihazı Doppler (; DE Hokanson, Inc; Belleveue, WA CW-1) bağlı olup,:; strain-gauge bir pletismografı (Bellevue, WA DE Hokanson, Inc AT-4) bağlanır. </li><li> Üst kol tıkanma manşon uygulanır, ve kol ayarlanabilir bir intravenöz kutup bağlı bir sapan bir bandaj ile kalp seviyesinde rahatça askıya alındı. Sistolik ve diyastolik kan basınçları ve kalp hızı karşısında kolu üzerine yerleştirilen bir Dinamap PROCARE 100 otomatik manşon (GE Healthcare, Piscataway, NJ) ile elde edilir. </li><li> Bileğine bir pediatrik manşet el akışını tıkamak için Hg 200 mm şişirilir. Üst kol manşet, 50 mm Hg kadar şişirilmiş 1,5 saniye sönük ve sonra önkol etrafında yerleştirilen strain gauge genişletilmesi yoluyla elde edilen her kol kan akımı ölçümü, hızla önce tekrar şişirilir. </li><li> Önkol kan akımı (FBF) tekrar dinlenmiş temel (FBF temel) ölçülür vet postocclusive hiperemi indüklenen maksimal vazodilatasyon (FBF max). Bazal kan akımı ölçümleri için, dört ardışık FBF eğrileri 30 sn (FBF tabanı) içinde elde edilmiştir. </li><li> Oklüzyon manşet sonra 10 dakika süreyle sistolik basıncının üzerinde 40 mm Hg şişirilir. Arteriyel oklüzyon (postocclusive reaktif hiperemi) salınımı takiben, dört ardışık FBF eğrileri akışının ilk 30 sn (FBF max) içinde elde edilmiştir. </li><li> Oranı FBF max / FBF tabanı dört FBF taban değerlerinin ortalaması ile dört FBF maksimum değerler ortalama bölerek, vazodilatasyon tahmini olarak hesaplanır. Maksimal vazodilatasyon (FVR maks) 5 Önkol vasküler direnç olarak hesaplanır FBF max bölü arter basıncı (OAB). Reaktif hiperemi sırasında FBF maksimum direkt olarak intra-arteryal infüzyonu asetilkolin maksimum, bir endotelyal-bağımlı damar genişletici sonra FBF ile ilgilidir. 6 </sup&gt; Buna göre, FBF max ve oranı FBF max / FBF baz endotel fonksiyonu non-invaziv önlemler. 6-8 ek olarak kabul edilir, FBF max ve FVR max hem direnç arter yapısal değişiklikler (artmış duvar / lümen oranı) yansıtmaktadır. 9 </li></ol>

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D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impaired+skincapillary+recruitment+in+essential+hypertension+is+caused+by+both+functional+and+structural+capillary+rarefaction.">Impaired skincapillary recruitment in essential hypertension is caused by both functional and structural capillary rarefaction.</a> Hypertension. 38, 238-242 (2001).</li><li>Carpentier, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+techniques+for+clinical+assessment+of+the+peripheral+microcirculation.">New techniques for clinical assessment of the peripheral microcirculation.</a> Drugs. 58, 17-22 (2001).</li><li>Shore, A. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Capillaroscopy+and+the+measurement+of+capillary+pressure.">Capillaroscopy and the measurement of capillary pressure.</a> Br. J. Clin. Pharmacol. 50, 501-513 (2000).</li></ol>

Yazarlar ifşa etmek bu çalışmanın yürütülmesi veya yayınlanmasına ilişkin ilgi hiçbir finansal çatışmalar var.

Kapillaroskopisi (kapiler mikroskopisi) kılcal yapısı, standart dışı bir ölçüsüdür. Bununla birlikte, şu anda, kılcal structure.Furthermore doğrudan değerlendirilmesi için standart yöntemler vardır, kılcal mikroskopi yaygın olarak yayınlanan çalışmalarında 12, 13, 10, 11, 14-18 büyük ve artan vücut içinde capillarydensity, doğrudan değerlendirilmesi için kullanılmıştır . Ayrıca, ön kol kan akımı, vasküler fonksiyonun bir köklü tedbir, 10, 11 ve BAE, damar ...

non-invasive assessment of microvascular and endothelial function

Yazarlar kardiyovasküler hastalık patogenezinde mikrovasküler disfonksiyon rolünü araştırmak için kapillaroskopisi ve önkol kan akım teknikleri kullanıldığında var. Kapillaroskopisi doğrudan mikrosirkülasyonu görselleştirmek için bir non-invaziv, nispeten ucuz yöntemdir. Yüzde kılcal işe maksimal kapiller yoğunluk (pasif venöz oklüzyon sırasında gözlenen) tarafından, postocclusive reaktif hiperemi (postocclusive reaktif hiperemi kapiller yoğunluk eksi başlangıçta kapiller yoğunluk) ile oluşturulan kapiller yoğunluk artışı bölünmesi ile tespit edilir. Yüzde kılcal (işlevsel aktif) perfüze tüm kılcal mevcut oranını temsil ve maksimal kapiller yoğunluğu postocclusive reaktif hiperemi kapiller yoğunluk bölünmesiyle hesaplanır perfüze. Yüzde kılcal işe alım ve yüzde Hem kılcal fonksiyonel kılcal sayısını yansıtmaktadır perfüze. Önkol kan akımı (FBF) tekniği provides endotel fonksiyonu non-invaziv önlemler alınır: oranı FBF max / FBF tabanı dört FBF temel değerlerinin ortalaması ile dört FBF maksimum değerler ortalama bölerek, vazodilatasyon tahmini olarak hesaplanır. Maksimal vazodilatasyon (FVR maks) Önkol vasküler direnç FBF max bölünmesiyle ortalama arter basıncı (OAB) olarak hesaplanmıştır. Kapillaroskopisi ve önkol teknikleri Her iki hasta kolayca kabul edilebilir ve hızlı bir şekilde öğrenilebilir. Mikrovasküler ve endotel fonksiyonu bu makalede açıklanan yöntemler kullanılarak elde edilen klinik hasta kardiyovasküler risk azaltma stratejileri gelecekte yarar olabilir ölçer. Biz yayınlanmış gibi olduğunu mikrovasküler ve endotel disfonksiyonu gösteren raporlar prehipertansiyon dahil hipertansiyonun ilk aşamalarında bulunan, mikrovasküler ve endotel fonksiyonu tedbirler zamanla erken tanımlanmasında yardımcı olabilir, Onun ölümcül sonuçları ile risk tabakalaşma ve son dönem vasküler patolojisi önlenmesi.

Normotansif ve hipertansif bireyler arasındaki mikrovasküler görünüş farklılıklar Şekiller 4 ve 5 karşılaştırılması ile kolayca görülür. Şekil 4, normotensif bireysel olarak iyi organize sıra düz kılcal tipik ağ gösterilmektedir. Buna karşın, Şekil 5, çökmüş, sarmal bir kapiller daha karıştırılmış deseni göstermektedir. Yazarlar kardiyovasküler hastalık patogenezinde mikrovasküler dis...

Watch this Scientific Journal Video about Non-invasive Assessment of Microvascular and Endothelial Function at JoVE.com

Non-invasive Assessment of Microvascular and Endothelial Function

Kapillaroskopisi doğrudan mikrosirkülasyonu görselleştirmek için bir non-invaziv, nispeten ucuz yöntemdir. Önkol kan akımı tekniği endotel fonksiyonu kabul non-invaziv önlemleri sağlar.

Mikrovasküler ve Endotel Fonksiyonu Noninvaziv Değerlendirilmesi

The authors have utilized capillaroscopy and forearm blood flow techniques to investigate the role of microvascular dysfunction in pathogenesis of ...

non-invasive-assessment-of-microvascular-and-endothelial-function

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

16.1K Görüntüleme.  Thomas Jefferson University. Kapillaroskopisi doğrudan mikrosirkülasyonu görselleştirmek için bir non-invaziv, nispeten ucuz yöntemdir. Önkol kan akımı tekniği endotel fonksiyonu kabul non-invaziv önlemleri sağlar.

Video: Non-invasive Assessment of Microvascular and Endothelial Function

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

Muscle strains are one of the most common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and physical exam alone, however the clinical presentation can vary greatly depending on the extent of injury, the patient's pain tolerance, etc. In patients with muscle injury or muscle disease, assessment of muscle damage is typically limited to clinical signs, such as tenderness, strength, range of motion, and more recently, imaging studies. Biological markers, such as serum creatine kinase levels, are typically elevated with muscle injury, but their levels do not always correlate with the loss of force production. This is even true of histological findings from animals, which provide a "direct measure" of damage, but do not account for all the loss of function. Some have argued that the most comprehensive measure of the overall health of the muscle in contractile force. Because muscle injury is a random event that occurs under a variety of biomechanical conditions, it is difficult to study. Here, we describe an in vivo animal model to measure torque and to produce a reliable muscle injury. We also describe our model for measurement of force from an isolated muscle in situ. Furthermore, we describe our small animal MRI procedure.

An in vivo Rodent Model of Contraction-induced Injury and Non-invasive Monitoring of Recovery

An in vivo animal model of injury is described. The method takes advantage of the subcutaneous position of the fibular nerve. Velocity, timing of muscle activation, and arc of motion are all pre-determined and synchronized using commercial software. Post injury changes are monitored in vivo using MR imaging/spectroscopy.

Bir In vivo</emDaralma indüklenen Sakatlık ve Kurtarma Non-invaziv İzleme> Kemirgen Model

Muscle strains are one of the most  common complaints treated by physicians. A muscle injury is typically diagnosed from the patient history and  physical ...

Tıp

Non-invasive Optical Measurement of Cerebral Metabolism and Hemodynamics in Infants

Perinatal brain injury remains a significant cause of infant mortality and morbidity, but there is not yet an effective bedside tool that can accurately screen for brain injury, monitor injury evolution, or assess response to therapy. The energy used by neurons is derived largely from tissue oxidative metabolism, and neural hyperactivity and cell death are reflected by corresponding changes in cerebral oxygen metabolism (CMRO2). Thus, measures of CMRO2 are reflective of neuronal viability and provide critical diagnostic information, making CMRO2 an ideal target for bedside measurement of brain health.
Brain-imaging techniques such as positron emission tomography (PET) and single-photon emission computed tomography (SPECT) yield measures of cerebral glucose and oxygen metabolism, but these techniques require the administration of radionucleotides, so they are used in only the most acute cases.
Continuous-wave near-infrared spectroscopy (CWNIRS) provides non-invasive and non-ionizing radiation measures of hemoglobin oxygen saturation (SO2) as a surrogate for cerebral oxygen consumption. However, SO2 is less than ideal as a surrogate for cerebral oxygen metabolism as it is influenced by both oxygen delivery and consumption. Furthermore, measurements of SO2 are not sensitive enough to detect brain injury hours after the insult 1,2, because oxygen consumption and delivery reach equilibrium after acute transients 3. We investigated the possibility of using more sophisticated NIRS optical methods to quantify cerebral oxygen metabolism at the bedside in healthy and brain-injured newborns. More specifically, we combined the frequency-domain NIRS (FDNIRS) measure of SO2 with the diffuse correlation spectroscopy (DCS) measure of blood flow index (CBFi) to yield an index of CMRO2 (CMRO2i) 4,5.
With the combined FDNIRS/DCS system we are able to quantify cerebral metabolism and hemodynamics. This represents an improvement over CWNIRS for detecting brain health, brain development, and response to therapy in neonates. Moreover, this method adheres to all neonatal intensive care unit (NICU) policies on infection control and institutional policies on laser safety. Future work will seek to integrate the two instruments to reduce acquisition time at the bedside and to implement real-time feedback on data quality to reduce the rate of data rejection.

We combined frequency-domain near-infrared spectroscopy measures of cerebral hemoglobin oxygenation with diffuse correlation spectroscopy measures of cerebral blood flow index to estimate an index of oxygen metabolism. We tested the utility of this measure as a bedside screening tool to evaluate the health and development of the newborn brain.

Bebeklerde Serebral Metabolizma ve Hemodinamiğin Noninvaziv Optik Ölçüm

Perinatal brain injury remains a significant cause of infant mortality  and morbidity, but there is not yet an effective bedside tool that can  accurately ...

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Mikrovasküler Fonksiyon ve Tadilat üzerine Arteriolar Ligasyonu etkisini değerlendirmek için mürin Spinotrapezius Modeli

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

A Methodological Approach to Non-invasive Assessments of Vascular Function and Morphology

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.

The present article describes the methodological considerations for several non-invasive assessments of vascular function and morphology that are commonly used in medical research to assess different stages of atherosclerosis.

Vasküler Fonksiyon ve Morfoloji Non-invaziv Değerlendirmelerine Bir Metodolojik Yaklaşım

The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis.  Damage to the endothelium may ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Canlı Fareler Seri endoskopik Görüntüleme kullanma Bağırsak Patolojilerde Yeni Therapeutics Etkinliğinin Non-invaziv Değerlendirme

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Ultrasound Assessment of Endothelial Function: A Technical Guideline of the Flow-mediated Dilation Test

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a pathological condition characterized mainly by a disruption in the balance between vasodilator and vasoconstrictor substances and is proposed to play an important role in the development of atherosclerotic cardiovascular disease. Therefore, a precise evaluation of endothelial function in humans represents an important tool that could help better understand the etiology of multiple cardio-centric pathologies.
Over the past twenty-five years, many methodological approaches have been developed to provide an assessment of endothelial function in humans. Introduced in 1989, the FMD test incorporates a forearm occlusion and subsequent reactive hyperemia that promotes nitric oxide production and vasodilation of the brachial artery. The FMD test is now the most widely utilized, non-invasive, ultrasonic assessment of endothelial function in humans and has been associated with future cardiovascular events.
Although the FMD test could have clinical utility, it is a physiological assessment that has inherited several confounding factors that need to be considered. This article describes a standardized protocol for determining FMD including the recommended methodology to help minimize the physiological and technical issues and improve the precision and reproducibility of the assessment.

The flow mediated dilation (FMD) test is the most commonly utilized, non-invasive, ultrasound assessment of endothelial function in humans. Although the FMD test has been related with the prediction of future cardiovascular disease and events, it is a physiological assessment with many inherent confounding factors that need to be considered.

Endotel Fonksiyon Ultrason Değerlendirmesi: Akış aracılı dilatasyon Testinin A Teknik Klavuzu

Cardiovascular disease is the primary cause of mortality and a major cause of disability worldwide. The dysfunction of the vascular endothelium is a ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. It is hypothesized that pro-atherogenic mediators released from fat tissues particularly in association with central/visceral adiposity may promote pathogenic vascular changes locally and systemically, and the notion that cardiovascular disease may be the consequence of adipose tissue dysfunction continues to evolve. Here, we describe a unique method of videomicroscopy that involves analysis of vasodilator and vasoconstrictor responses of intact small human arterioles removed from the adipose depot of living human subjects. Videomicroscopy is used to examine functional properties of isolated microvessels in response to pharmacological or physiological stimuli using a pressured system that mimics in vivo conditions. The technique is a useful approach to gain understanding of the pathophysiology and molecular mechanisms that contribute to vascular dysfunction locally within the adipose tissue milieu. Moreover, abnormalities in the adipose tissue microvasculature have also been linked with systemic diseases. We applied this technique to examine depot-specific vascular responses in obese subjects. We assessed endothelium-dependent vasodilation to both increased flow and acetylcholine in adipose arterioles (50 - 350 &#181;m internal diameter, 2 - 3 mm in length) isolated from two different adipose depots during bariatric surgery from the same individual. We demonstrated that arterioles from visceral fat exhibit impaired endothelium-dependent vasodilation compared to vessels isolated from the subcutaneous depot. The findings suggest that the visceral microenvironment is associated with vascular endothelial dysfunction which may be relevant to clinical observation linking increased visceral adiposity to systemic disease mechanisms. The videomicroscopy technique can be used to examine vascular phenotypes from different fat depots as well as compare findings across individuals with different degrees of obesity and metabolic dysfunction. The method can also be used to examine vascular responses longitudinally in response to clinical interventions.

Videomicroscopy systems are used to examine functional properties of isolated adipose tissue arterioles in response to physiological and pharmacological stimuli. This technique can be used to examine microvascular phenotypes in different adipose tissue domains in obese humans.

İnsan yağ dokusu mikrovasküler işlevini kullanarak Videomicroscopy değerlendirilmesi

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Assessment of Kidney Function in Mouse Models of Glomerular Disease

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function in diabetic nephropathy and podocyte-specific VEGF-A knock-out mice; therefore, this protocol describes the full kidney work-up used in our lab to assess these mouse models of glomerular disease, enabling a vast amount of information regarding kidney and glomerular function to be obtained from a single mouse. In comparison to alternative methods presented in the literature to assess glomerular function, the use of the method outlined in this paper enables the glomerular phenotype to be fully evaluated from multiple aspects. By using this method, the researcher can determine the kidney phenotype of the model and assess the mechanism as to why the phenotype develops. This vital information on the mechanism of disease is required when examining potential therapeutic avenues in these models. The methods allow for detailed functional assessment of the glomerular filtration barrier through measurement of the urinary albumin creatinine ratio and individual glomerular water permeability, as well as both structural and ultra-structural examination using the Periodic Acid Schiff stain and electron microscopy. Furthermore, analysis of the genes dysregulated at the mRNA and protein level enables mechanistic analysis of glomerular function. This protocol outlines the generic but adaptable methods that can be applied to all mouse models of glomerular disease.

This protocol describes a full kidney work-up that should be carried out in mouse models of glomerular disease. The methods allow for detailed functional, structural, and mechanistic analysis of glomerular function, which can be applied to all mouse models of glomerular disease.

Fare modelleri Glomerular hastalığı böbrek fonksiyonlarının değerlendirilmesi

The use of murine models to mimic human kidney disease is becoming increasingly common. Our research is focused on the assessment of glomerular function ...

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively affect skeletal muscle. This can result in a loss of muscle mass (atrophy) and/or loss of muscle quality (reduced force per unit of muscle mass), both of which are prevalent in chronic disease, muscle-specific disease, immobilization, and aging (sarcopenia). Skeletal muscle function in animals can be evaluated by a range of different tests. All tests have limitations related to the physiological testing environment, and the selection of a specific test often depends on the nature of the experiments. Here, we describe an in vivo, non-invasive technique involving a helpful and easy assessment of force frequency-curve (FFC) in mice that can be performed on the same animal over time. This permits monitoring of disease progression and/or efficacy&#160;of a potential therapeutic treatment.

Measurement of rodent skeletal muscle contractile function is a useful tool that can be used to track disease progression as well as efficacy of therapeutic intervention. We describe here the non-invasive, in vivo assessment of the dorsiflexor muscles that can be repeated over time in the same mouse.

Non-invaziv Dorsiflexor kas fonksiyonu farelerde değerlendirilmesi

Assessment of skeletal muscle contractile function is an important measurement for both clinical and research purposes. Numerous conditions can negatively ...

A Computerized Functional Skills Assessment and Training Program Targeting Technology Based Everyday Functional Skills

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a computerized functional skills training program that was paired in half of the participants with a commercially available cognitive training (CCT) program.
Non-impaired older individuals (NC) aged 60+ (n=45) and similarly aged individuals with mild cognitive impairment (MCI; n=50) were randomized to receive 12 weeks of twice-weekly computerized functional skills training (CFST) or 12 weeks of twice-weekly sessions split between CCT and CFST. Skills trained were use of an ATM; internet banking; ticket kiosk; telephone and internet prescription refill; medication management; and internet shopping. As with previous functional capacity assessments, we focus on completion time for each simulation.
51 participants completed the training program, either by mastering all 6 tasks (34) or completing 12 weeks of training. 44 more participants completed 4 or more training sessions so they were also analyzed for improvement up to their last training session. Completion time for all 6 tests significantly improved from the baseline assessment to the final training session in both groups of participants (all p&#60;0.001 with an average improvement in task completion time of 45%). Further, there was no differential improvement in MCI and NC in the 6 tests from baseline to end of training (all t&#60;1.66, all p&#62;0.12). Finally, combined CCT plus CFST did not differ from CSFT alone on any of the percent-change score measures (all t&#60;1.64, all p&#62;0.11).
Both NC and MCI groups evidenced substantial improvements in performance. CCT supplementation led to similar functional gains with half as many training sessions. The NC participants proceeded through the training fairly rapidly even without CCT supplementation; MCI participants required more training but learned equivalently. These findings suggest that even in cases with memory impairments, functional skills can be efficiently learned with training.

This training protocol uses computerized training to teach technology-related everyday functional skills. These skills include financial skills, travel and transit, as well as medication management.

Mikrovasküler ve Endotel Fonksiyonu Noninvaziv Değerlendirilmesi

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