This project was supported by Grant Numbers HL96593 from NIH and D56HP20783 from HRSA/ HHS. Its contents are solely the responsibility of the authors and do not necessarily represent the official views of the NIH or HRSA / HHS.

Remarque: Le processus d&#39;acquisition pour obtenir des images capillaires a déjà été publié et est réalisée en utilisant un appareil photo numérique avec un programme informatique d&#39;acquisition et d&#39;analyse d&#39;image correspondante 11 21 Ce laboratoire utilise des images fixes pour l&#39;analyse, pas les vidéos, ce qui simplifie l&#39;acquisition d&#39;image pour l&#39;analyse.. Ce qui suit décrit la nouvelle technique pour quantifier les capillaires à partir des images. 1. Amélioration de l&#39;image processus <ol><li> Obtenir des images numériques avec un appareil photo numérique monochrome. Calibrer les images à un objet de taille connue en prenant une photo d&#39;un objet de longueur connue comme une règle avec l&#39;appareil photo. Ce processus permet au programme d&#39;ordinateur pour mesurer et compter capillaires précision après traitement. Idéalement pour la plus haute précision, un réticule (pièce scientifiquement fabriqué en verre avec une règle gravée en elle) devrait être utilisé. Mesurer le nombre de pixels dans un carré de 1 mmboîte à l&#39;aide d&#39;un programme informatique. Remarque: La clé de la reproductibilité et de la normalisation de ce protocole dépend largement de placement correct de la boîte de 1 mm 2 qui est compté. </li><li> Utilisez des outils d&#39;amélioration de contraste pour assombrir les capillaires et éclaircir le fond, ce qui permettra de maximiser la visualisation des capillaires. Différenciation initiale de capillaires de fond est important pour le bon recadrage de l&#39;image dans les étapes ultérieures. Pour ce faire, réglage de l&#39;image en utilisant un Pour ce faire, cliquez sur &quot;meilleure histogramme ajustement.&quot;: La capture, de l&#39;intensité, histogramme de l&#39;image, meilleur ajustement. </li><li> Cadrer une région d&#39;intérêt (ROI) pour capillaire comptage comme une nouvelle image (sélectionner, régions d&#39;intérêt). Utilisez une boîte de 1 mm 2, qui a été déterminé par l&#39;étalonnage des images à 530 pixels équivalant à 1 mm. vérifier que l&#39;image recadrée place au sommet des boucles capillaires tout en haut de l&#39;image. </li><li> Aplatir l&#39;image de sorte que tous les futurs ajustements d&#39;image seront appliqués uniformémentl&#39;image. Pour ce faire, cliquez sur: onglet processus, filtres 2D, aplatir, l&#39;intensité de BG &quot;brillant&quot;, la largeur de l&#39;objet à &quot;75,&quot; applique. </li><li> Augmenter le contraste de l&#39;image afin capillaires sont visualisés au maximum. Pour ce faire, cliquez sur: ajuster onglet, affichage, augmenter le contraste à 75. </li><li> Dépoussiérer l&#39;image pour lisser les bords des capillaires en cliquant sur l&#39;onglet processus, filtres 2D, déparasitage, la taille du noyau 7 x 7, appliquer. </li><li> Finaliser le contraste de l&#39;image afin capillaires sont le noir et le fond est blanc. Effectuez cette étape en ajustant l&#39;histogramme pour le modèle &quot;meilleur ajustement&quot;. Note: S&#39;il vous plaît se référer à la figure 2 en résultats représentatifs pour un exemple de ce que l&#39;image traitée devrait ressembler suivant ces étapes. </li></ol> 2. Exécution comtes capillaires / quantification capillaire Densité <ol><li> Sur chaque image, sélectionner manuellement une partie d&#39;un capillaire bien définie en utilisant la featur &quot;objet cible&quot;e d&#39;être reconnu comme objets à compter par le programme. Ensuite, sélectionnez une petite partie du fond, en utilisant la fonction «de fond», comme une référence à des zones qui doivent être ignorés par l&#39;algorithme de comptage. Remarque: La combinaison de ces faits saillants provoque tous les capillaires à être mis en évidence en faisant abstraction du bruit de fond. La quantification (comptage) protocole utilise les fonctions informatiques de différencier parties de l&#39;image basée sur la couleur et la morphologie. </li><li> Utilisez la fonction de comptage pour compter instantanément tous les capillaires dans l&#39;image avec l&#39;équipement d&#39;imagerie. Réglez le diamètre minimum d&#39;objets comptés à 5 pixels afin d&#39;éviter de compter le bruit de fond que les capillaires. </li><li> Pour chaque individu, compter le moyenne de 3 - 4 images afin d&#39;obtenir une évaluation plus fiable. Note: S&#39;il vous plaît se référer à la figure 3 dans Les résultats représentatifs pour un exemple de ce que l&#39;image devrait ressembler au cours de la procédure de comptage. </li></ol&gt; 3. Création et utilisation de macros pour automatiser le traitement de l&#39;image Remarque: Pour gagner du temps, les macros peuvent être créées afin d&#39;effectuer automatiquement une séquence spécifique des processus sur une ou plusieurs images. Ces séquences peuvent être personnalisés afin de rendre les modifications de l&#39;image rapide. En substance, ces macros se rappeler comment les images sont traitées, et d&#39;effectuer toutes les étapes rapidement et sans intervention de l&#39;utilisateur. Performing compte sur 12 images capillaires prend ce laboratoire entre 20 à 30 min avec les macros (2 à 3 min par image), par opposition à environ 8 min par image sans les macros. Par conséquent en utilisant les macros est de 3 à 5 fois plus efficace que le traitement manuellement chaque image individuelle. <ol><li> Afin de créer une macro, sélectionnez &quot;Enregistrer une macro&quot; et sur une image effectuent les étapes et les processus désirés, comme décrit dans les étapes 1 et 2 ci-dessus. Nom de la macro sur la base duquel le traitement d&#39;image étapes ont été réalisées pour référence future. Lorsque using la macro sur les images futures, cliquez simplement sur &quot;macro run&quot; et le programme va appliquer automatiquement les améliorations enregistrées à l&#39;image (s) souhaitée. Note: Ce laboratoire utilise une macro pour effectuer tous, mais l&#39;une des étapes de la Section 1 des méthodes en quelques secondes. La seule étape qui nécessite une entrée de l&#39;utilisateur est de choisir où pour recadrer l&#39;image, l&#39;étape 1.2. </li></ol>

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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=Computerized+nailfold+video+capillaroscopy--a+new+tool+for+assessment+of+Raynaud's+phenomenon.">Computerized nailfold video capillaroscopy--a new tool for assessment of Raynaud's phenomenon.</a> J Rheumatol. 32, 841-848 (2005).</li><li>Neubauer-Geryk, J., 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=Decreased+reactivity+of+skin+microcirculation+in+response+to+L-arginine+in+later-onset+type+1+diabetes.">Decreased reactivity of skin microcirculation in response to L-arginine in later-onset type 1 diabetes.</a> Diabetes Care. 36, 950-956 (2013).</li><li>Pazos-Moura, C. C., Moura, E. G., Bouskela, E., Torres-Filho, I. P., Breitenbach, M. 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B., Tibirica, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antihypertensive+treatment+improves+microvascular+rarefaction+and+reactivity+in+low-risk+hypertensive+individuals.">Antihypertensive treatment improves microvascular rarefaction and reactivity in low-risk hypertensive individuals.</a> Microcirculation. 20, 703-716 (2013).</li><li>Cheng, C., Diamond, J. J., Falkner, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+capillary+rarefaction+in+mild+blood+pressure+elevation.">Functional capillary rarefaction in mild blood pressure elevation.</a> Clinical and Translational Science. 1, 75-79 (2008).</li><li>Debbabi, H., Bonnin, P., Levy, B. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+blood+pressure+control+with+perindopril/indapamide+on+the+microcirculation+in+hypertensive+patients.">Effects of blood pressure control with perindopril/indapamide on the microcirculation in hypertensive patients.</a> Am J Hypertens. 23, 1136-1143 (2010).</li><li>He, F. J., Marciniak, M., Markandu, N. D., Antonios, T. F., MacGregor, G. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+modest+salt+reduction+on+skin+capillary+rarefaction+in+white,+black,+and+Asian+individuals+with+mild+hypertension.">Effect of modest salt reduction on skin capillary rarefaction in white, black, and Asian individuals with mild hypertension.</a> Hypertension. 56, 253-259 (2010).</li><li>Murray, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+measurement+location+on+reliability+of+quantitative+nailfold+videocapillaroscopy+in+patients+with+SSc.">The influence of measurement location on reliability of quantitative nailfold videocapillaroscopy in patients with SSc.</a> Rheumatology (Oxford). 51, 1323-1330 (2012).</li><li>Ingegnoli, F., 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=Feasibility+of+different+capillaroscopic+measures+for+identifying+nailfold+microvascular+alterations.">Feasibility of different capillaroscopic measures for identifying nailfold microvascular alterations.</a> Semin Arthritis Rheum. 38, 289-295 (2009).</li><li>Debbabi, 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=Increased+skin+capillary+density+in+treated+essential+hypertensive+patients.">Increased skin capillary density in treated essential hypertensive patients.</a> Am J Hypertens. 19, 477-483 (2006).</li><li>Serne, E. 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=Impaired+skin+capillary+recruitment+in+essential+hypertension+is+caused+by+both+functional+and+structural+capillary+rarefaction.">Impaired skin capillary recruitment in essential hypertension is caused by both functional and structural capillary rarefaction.</a> Hypertension. 38, 238-242 (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><li>Rieder, M. J., O'Drobinak, D. M., Greene, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+computerized+method+for+determination+of+microvascular+density.">A computerized method for determination of microvascular density.</a> Microvasc Res. 49, 180-189 (1995).</li><li>Vermeulen, P. 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The authors have no conflicts of interest.

Capillaroscopie périunguéale montre promesse comme un outil clinique utile à l&#39;avenir pour diverses oncologie, cardiovasculaire, et les applications de maladies rhumatologiques. Le processus d&#39;acquisition d&#39;image est assez uniforme entre les chercheurs, mais il ya actuellement plusieurs méthodes pour le traitement et l&#39;analyse d&#39;images. Méthodes comprennent actuellement comptages manuels et informatisés capillaires. Des comptages manuels sont problématiques car ils prennent du temps, et sous r...

Imagerie microvasculaire est un domaine en pleine expansion avec de nombreuses applications cliniques potentielles. 2 Par exemple, les oncologues utilisent l&#39;imagerie des microvaisseaux pour déterminer l&#39;étendue de l&#39;angiogenèse tumorale, ce qui donne de précieuses informations concernant l&#39;état de la tumeur et la perspicacité dans les options de traitement possibles. 3 4 Cependant, périunguéale capillaroscopie est peut-être le coût forme la plus efficace et largement applicable de l&#39;imagerie microvasculaire. Les chercheurs utilisent la vidéo périunguéale capillaroscopie pour étudier les débits sanguins et d&#39;enquêter sur la morphologie capillaire. 5 6 fois la vidéo et les images fixes, capillaroscopie périunguéale sont adjoints aux soins pour le diagnostic et le traitement des maladies du tissu conjonctif phénomène et divers de Raynaud comme la sclérose systémique. 2 Capillaroscopie périunguéale a diverses applications cardiovasculaires potentiels ainsi. La recherche actuelle en utilisant périunguéale capillaroscopie suggèreque diabète sucré de type 1 et de type 2 patients présentent une prévalence élevée de la morphologie capillaire anormale, mais ont des densités capillaires inchangé par rapport aux personnes non-diabétiques. 7-8 capillaroscopie a également été étudié expérimentalement dans l&#39;hypertension. Structurelle raréfaction capillaire conduisant à une densité capillaire réduite a été démontrée chez les personnes hypertendues par rapport aux individus non-hypertendus. 9-10 Contrairement à ces patients hypertendus âgés (âge moyen 40 ans et plus) qui présentent la raréfaction structurelle, des recherches plus récentes ont démontré que patients hypertendus plus jeunes (moyenne âge de moins de 40 ans) ont raréfaction fonctionnel sans raréfaction structurelle. 11 Ceci suggère que la raréfaction fonctionnelle se produit avant et peut évoluer au fil du temps à la raréfaction structurelle. Fait intéressant, les patients hypertendus traités avec des médicaments antihypertenseurs spécifiques tels que perindopril / indapamide affiché normedensité al capillaire et la fonction endothéliale après le traitement, tandis que ceux traités avec ACE (angiotensin l&#39;enzyme de conversion) des inhibiteurs ou des diurétiques maintenu une densité capillaire faible malgré le contrôle de la pression artérielle comparable. 12 Ceci suggère que certains médicaments anti-hypertenseurs peuvent normaliser la densité capillaire en inversant le tube capillaire raréfaction provoquée par l&#39;hypertension. En outre, d&#39;autres chercheurs ont montré que la réduction de la consommation de sel conduit à l&#39;inversion de deux raréfaction capillaire fonctionnelle et structurale chez les personnes hypertendues 13. Malgré les diverses applications cliniques potentielles de cette technologie, il ya peu de standardisation de la technique pour quantifier images de densité capillaire. 2 A ce jour, les chercheurs ont constaté que les résultats de la densité capillaire sont reproductibles à la fois intra-observateur et inter-observateur perspective que si l&#39;exacte même zone est compté à chaque fois. 15 1,14 </sup&gt; Fait à noter, les chercheurs précédents ont largement effectué chiffres capillaires manuellement en utilisant l&#39;œil nu, 9 16 17 18 qui est un processus lent et subjective. , Sur la base des algorithmes informatiques standardisés pour la quantification des images capillaires offrent théoriquement analyse de données plus efficace et reproductible avec moins de subjectivité, de faciliter les applications cliniques de capillaroscopie. Certains chercheurs ont en effet utilisé des programmes informatiques pour quantifier les données de périunguéale photos capillaroscopiques. 1,6 19 20 Toutefois, une seule publication à ce jour décrit la fiabilité d&#39;un logiciel complexe disponibles pour quantifier les données de capillaroscopie, 1 et ce programme est compliqué comme précédemment noté ci-dessus par l&#39;obligation de compter le même champ visuel exacte. Ici, nous présentons, un protocole simple fiables pour quantifier capillaires en utilisant un algorithme standardisé qui permet del&#39;utilisation de plusieurs champs visuels. L&#39;utilisation de plusieurs champs visuels non seulement simplifie la procédure, mais permet également à l&#39;évaluation de la variation biologique normale du nombre de capillaires. Le but de cette étude est de décrire un algorithme basé sur ordinateur reproductible et efficace qui normalise le processus de quantification capillaire. Bien que ces méthodes ne sont pas entièrement automatisés, ils nécessitent très peu d&#39;entrée de l&#39;utilisateur, et fournissent une quantification fiable et rapide des photos.

<table>
	<tbody>
		<tr>
			<td class="xl65">Image-Pro Premier</td>
			<td class="xl65">Media Cybernetics, Inc</td>
			<td align="right" class="xl63">9.1</td>
			<td class="xl63">Image processing software</td>
		</tr>
	</tbody>
</table>

a reproducible computerized method for quantitation of capillary density using nailfold capillaroscopy

Capillaroscopie est une méthode non-invasive, efficace, relativement peu coûteux et facile à apprendre pour visualiser directement la microcirculation. La technique de capillaroscopie peut donner un aperçu de microvasculaire la santé d&#39;un patient, conduisant à une variété de dermatologique potentiellement précieux, ophtalmologique, les applications cliniques rhumatologiques et cardiovasculaires. En outre, la croissance tumorale peut être dépendante de l&#39;angiogenèse, qui peut être quantifiée par la mesure de la densité des microvaisseaux dans la tumeur. Cependant, il est actuellement peu ou pas de standardisation des techniques, et une seule publication à ce jour signale la fiabilité d&#39;un algorithmes actuellement disponibles, complexes basés ordinateur pour quantifier des données de capillaroscopie. 1 Ce document décrit un nouveau, plus simple, fiable, capillaire standardisé comptage algorithme pour quantifier les données de capillaroscopie périunguéale. Un algorithme simple, reproductible informatisée capillaroscopie comme cela faciliterait plusutilisation généralisée de la technique entre les chercheurs et les cliniciens. De nombreux chercheurs analysent actuellement les images capillaroscopie à la main, la promotion de la fatigue de l&#39;utilisateur et de la subjectivité des résultats. Ce document décrit un roman, facile à utiliser automatisé algorithme de traitement d&#39;image en plus, un algorithme de comptage semi-automatisé reproductible. Cet algorithme permet l&#39;analyse d&#39;images en quelques minutes tout en réduisant la subjectivité; seulement une quantité minimale de temps de formation (dans notre expérience, moins de 1 h) est nécessaire pour apprendre la technique.

L&#39;objectif de cette procédure de traitement d&#39;image est de différencier les capillaires de l&#39;image d&#39;arrière-plan de sorte qu&#39;ils peuvent être quantifiés avec précision. Tant de traitement d&#39;image incomplète et traitement d&#39;image excessive nuisent à la capacité du programme à quantifier les capillaires. Comme on le voit sur ​​la figure 3, le traitement d&#39;image effectue les capillaires incomplet difficile à distinguer de l&#39;arrière-plan....

Watch this Scientific Journal Video about A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy at JoVE.com

A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm.

Une méthode informatisée reproductible pour la quantification de la densité capillaire utilisant périunguéale capillaroscopie

Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy to learn methodology for directly visualizing the microcirculation. The ...

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8.9K vues.  Thomas Jefferson University. Capillaroscopy is a non-invasive, efficient, relatively inexpensive and easy-to-learn methodology for directly visualizing capillaries in the microcirculation. However, only one publication to date describes the reliability of a complex software program available for quantitating capillaroscopy data. Here, we present a simple, reliable protocol for quantitating capillaries using a standardized algorithm. 

Vidéo: A Reproducible Computerized Method for Quantitation of Capillary Density using Nailfold Capillaroscopy

Photothrombotic Ischemia: A Minimally Invasive and Reproducible Photochemical Cortical Lesion Model for Mouse Stroke Studies

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected light-sensitive dye. Following illumination, the dye is activated and produces singlet oxygen that damages components of endothelial cell membranes, with subsequent platelet aggregation and thrombi formation, which eventually determines the interruption of local blood flow. This approach, initially proposed by Rosenblum and El-Sabban in 1977, was later improved by Watson in 1985 in rat brain and set the basis of the current model. Also, the increased availability of transgenic mouse lines further contributed to raise the interest on the photothrombosis model. Briefly, a photosensitive dye (Rose Bengal) is injected intraperitoneally and enters the blood stream. When illuminated by a cold light source, the dye becomes activated and induces endothelial damage with platelet activation and thrombosis, resulting in local blood flow interruption. The light source can be applied on the intact skull with no need of craniotomy, which allows targeting of any cortical area of interest in a reproducible and non-invasive way. The mouse is then sutured and allowed to wake up. The evaluation of ischemic damage can be quickly accomplished by triphenyl-tetrazolium chloride or cresyl violet staining. This technique produces infarction of small size and well-delimited boundaries, which is highly advantageous for precise cell characterization or functional studies. Furthermore, it is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Photothrombosis is a quick, minimally-invasive technique for inducing small and well-delimited infarction in areas of interest in highly reproducible manner. It is particularly suitable for studying cellular and molecular responses underlying brain plasticity in transgenic mice.

Ischémie Photothrombotic: A photochimique modèle de lésion corticale minimalement invasive et reproductible pour les études de course de souris

The photothrombotic stroke model aims to induce an ischemic damage within a given cortical area by means of photo-activation of a previously injected ...

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Médecine

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

Génération et quantification 3-Dimensional des lésions artérielles chez la souris utilisant la tomographie optique de projection

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic conditions. As part of a diagnostic assessment, the early detection of postural deficits is important so that appropriate and targeted interventions can be prescribed. The Sensory Organization Test (SOT) on the EquiTest determines an individual&#39;s use of the sensory systems (somatosensory, visual, and vestibular) that are responsible for postural control. Somatosensory and visual input are altered by the calibrated sway-referenced support surface and visual surround, which move in the anterior-posterior direction in response to the individual&#39;s postural sway. This creates a conflicting sensory experience. The Motor Control Test (MCT) challenges postural control by creating unexpected postural disturbances in the form of backwards and forwards translations. The translations are graded in magnitude and the time to recover from the perturbation is computed.
Intermittent claudication, the most common symptom of peripheral arterial disease, is characterized by a cramping pain in the lower limbs and caused by muscle ischemia secondary to reduced blood flow to working muscles during physical exertion. Claudicants often display poor balance, making them susceptible to falls and activity avoidance. The Ankle Brachial Pressure Index (ABPI) is a noninvasive method for indicating the presence of peripheral arterial disease and intermittent claudication, a common symptom in the lower extremities. ABPI is measured as the highest systolic pressure from either the dorsalis pedis or posterior tibial artery divided by the highest brachial artery systolic pressure from either arm. This paper will focus on the use of computerized dynamic posturography in the assessment of balance in claudicants.

Individuals with intermittent claudication exhibit poor balance compared to healthy controls. Computerized dynamic posturography is an objective method for measuring an individual's postural responses to balance disturbances. This provides an objective reflection of the person&#x2019;s ability to respond to situations under which sensory stimuli are altered and unexpected perturbations occur.

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

Quantification du commerce intra-péritonéale Cancer de l&#39;ovaire Métastase

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Method of Direct Segmental Intra-hepatic Delivery Using a Rat Liver Hilar Clamp Model

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ischemia/reperfusion (I/R) injury with myriad negative consequences. Potential I/R injury in marginal organs destined for liver transplantation contributes to the current donor shortage secondary to a decreased organ utilization rate. A significant need exists to explore hepatic I/R injury in order to mediate its impact on graft function in transplantation. Rat liver hilar clamp models are used to investigate the impact of different molecules on hepatic I/R injury. Depending on the model, these molecules have been delivered using inhalation, epidural infusion, intraperitoneal injection, intravenous administration or injection into the peripheral superior mesenteric vein. A rat liver hilar clamp model has been developed for use in studying the impact of pharmacologic molecules in ameliorating I/R injury. The described model for rat liver hilar clamp includes direct cannulation of the portal supply to the ischemic hepatic segment via a side branch of the portal vein, allowing for direct segmental hepatic delivery. Our approach is to induce ischemia in the left lateral and median lobes for 60 min, during which time the substance under study is infused. In this case, pegylated-superoxide dismutase (PEG-SOD), a free radical scavenger, is infused directly into the ischemic segment. This series of experiments demonstrates that infusion of PEG-SOD is protective against hepatic I/R injury. Advantages of this approach include direct injection of the molecule into the ischemic segment with consequent decrease in volume of distribution and reduction in systemic side effects.

A unique rat liver hilar clamp model was developed for studying the impact of pharmacologic molecules in ameliorating ischemia-reperfusion injury. This model includes direct cannulation of the portal supply to the ischemic liver segment via a branch of the portal vein, allowing for direct hepatic delivery.

Méthode de Direct segmentaire livraison intra-hépatique L&#39;utilisation d&#39;un modèle hilaire de foie de rat Clamp

Major hepatic surgery with inflow occlusion, and liver transplantation, necessitate a period of warm ischemia, and a period of reperfusion leading to ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

Reproductible Arterial dénudation blessures par infrarénale de l&#39;aorte abdominale de serrage dans un modèle murin

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Method for Quantifying Upper Limb Performance in Daily Life Using Accelerometers

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. It has become important to measure a person's activities in daily life, and not just measure their capacity for activity in the structured environment of a clinic or laboratory. A wearable sensor that is now enabling measurement of daily movement is the accelerometer. Accelerometers are commercially-available devices resembling large wrist watches that can be worn throughout the day. Data from accelerometers can quantify how the limbs are engaged to perform activities in peoples' homes and communities. This report describes a methodology to collect accelerometry data and turn it into clinically-relevant information. First, data are collected by having the participant wear two accelerometers (one on each wrist) for 24 h or longer. The accelerometry data are then downloaded and processed to produce four different variables that describe key aspects of upper limb activity in daily life: hours of use, use ratio, magnitude ratio, and the bilateral magnitude. Density plots can be constructed that visually represent the data from the 24 h wearing period. The variables and their resultant density plots are highly consistent in neurologically-intact, community-dwelling adults. This striking consistency makes them a useful tool for determining if upper limb daily performance is different from normal. This methodology is appropriate for research studies investigating upper limb dysfunction and interventions designed to improve upper limb performance in daily life in people with stroke and other patient populations. Because of its relative simplicity, it may not be long before it is also incorporated in clinical neurorehabilitation practice.

This protocol describes a method to quantify upper limb performance in daily life using wrist-worn accelerometers.

Une méthode de quantification des performances du membre supérieur dans la vie quotidienne utilisant des accéléromètres

A key reason for referral to rehabilitation services after stroke and other neurological conditions is to improve one's ability to function in daily life. ...

Ablation of Ischemic Ventricular Tachycardia Using a Multipolar Catheter and 3-dimensional Mapping System for High-density Electro-anatomical Reconstruction

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars represent zones of slow conduction that allow the occurrence and maintenance of reentrant circuits. Catheter ablation enables substrate modification of those low voltage areas and thus can help to alter the scar tissue in such a way that arrhythmias cannot appear anymore. Hospitalizations of concerned patients decrease, quality of life and outcome rise. Consequently, VT ablation represents a growing field in electrophysiology, especially for patients with endocardial scars in ischemic heart disease after myocardial infarction. However, ablation of ventricular tachycardia remains one of the most challenging procedures in the electrophysiology lab. Precise scar definition and localization of abnormal potentials are critical for ablation success. The following manuscript describes the use of a multipolar mapping catheter and 3-dimensional (3D) mapping system to create a high density electro-anatomical map of the left ventricle including a precise scar representation as well as mapping of fractionated and late potentials in order to allow a highly accurate substrate modification.

With the following protocol, we provide an approach to Ventricular Tachycardia (VT) ablation using high density mapping with a multipolar catheter and 3D mapping system enhancing the success of the procedure.

Ablation de la tachycardie ventriculaire ischémique en utilisant un cathéter multipolaire et le système de cartographie 3D pour la Reconstruction de Electro-anatomique haute densitée

Ventricular tachycardia (VT) in patients with ischemic cardiomyopathy mainly results from endocardial scars after myocardial infarction; those scars ...

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.

Un programme informatisé d’évaluation et de formation des compétences fonctionnelles ciblant la technologie basée sur les compétences fonctionnelles quotidiennes

Today, many functional skills are technology-based, so development of a technology-based training program has broad importance. Here we present a ...

A Reproducible Intensive Care Unit-Oriented Endotoxin Model in Rats

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ranges between 20 and 30%. Novel treatment approaches in order to reduce sepsis-related multiorgan failure and death are urgently needed. Robust animal models allow for one or multiple treatment approaches as well as for testing their effect on physiological and molecular parameters. In this article, a simple animal model is presented.
First, general anesthesia is induced in animals either with the use of volatile or by intraperitoneal anesthesia. After placement of an intravenous catheter (tail vein), tracheostomy, and insertion of an intraarterial catheter (tail artery), mechanical ventilation is started. Baseline values of mean arterial blood pressure, arterial blood oxygen saturation, and heart rate are recorded.
The injection of lipopolysaccharides (1 milligram/kilogram body weight) dissolved in phosphate-buffered saline induces a strong and reproducible inflammatory response via the toll-like receptor 4. Fluid corrections as well as the application of norepinephrine are performed based on well-established protocols.
The animal model presented in this article is easy to learn and strongly oriented towards clinical sepsis treatment in an intensive care unit with sedation, mechanical ventilation, continuous blood pressure monitoring, and repetitive blood sampling. Also, the model is reliable, allowing for reproducible data with a limited number of animals in accordance with the 3R (reduce, replace, refine) principles of animal research. While animal experiments in sepsis research cannot easily replaced, repetitive measurements allow for a reduction of animals and keeping septic animals anesthetized diminishes suffering.

Here, we present a reproducible intensive care unit-oriented endotoxin model in rats.

Un modèle d’endotoxine reproductible orienté unité de soins intensifs chez le rat

Sepsis and septic shock remain the leading cause of death in intensive care units. Despite significant improvements in sepsis management, mortality still ...