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.

Hinweis: Die Übernahme Verfahren zur Gewinnung von kapillaren Bilder vorher veröffentlicht worden und wird mit einem noch Digitalkamera mit einer entsprechenden Bildaufnahme und Analyse Computerprogramm erreicht 11 21 Diese Übung nutzt Standbilder für die Analyse, keine Videos, die Vereinfachung der Bildaufnahme für die Analyse.. Im folgenden wird die neue Technik für die Quantifizierung der Kapillaren aus den Bildern. 1. Bildoptimierung Prozess <ol><li> Erhalten Digitalbilder mit einer monochromen Digitalkamera. Kalibrieren Sie die Bilder zu einem Objekt bekannter Größe durch die ein Bild eines Objekts mit bekannter Länge wie ein Lineal mit der Kamera. Dieser Prozess ermöglicht es dem Computerprogramm zu messen und zu zählen, Kapillaren exakt nach der Verarbeitung. Im Idealfall für höchste Präzision, sollte ein Fadenkreuz (wissenschaftlich hergestellten Stück Glas mit einem Lineal hinein geätzt) verwendet werden. Ermittlung der Zahl der Pixel in einem 1 mm QuadratKasten mit einem Computerprogramm. Anmerkung: Der Schlüssel für die Reproduzierbarkeit und Standardisierung dieses Protokoll, hängt weitgehend von der richtigen Plazierung des 1 mm 2 Feld gezählt wird. </li><li> Verwenden Enhancement Tools, um die Kapillaren zu verdunkeln und erhellen den Hintergrund, die Visualisierung der Kapillaren maximiert. Initial Differenzierung von Kapillaren aus Hintergrund ist für eine gute Abschneiden des Bildes in späteren Schritten. Tun Sie dies, indem Sie das Bild mit einem Klicken Sie dazu auf zu tun &quot;best fit Histogramm.&quot;: Erfassung, Intensität, Histogramm, beste Passform. </li><li> Zuschneiden von Region of Interest (ROI) für die Kapillar-Zählung als ein neues Bild (Wahlen, Regions of Interest). Verwenden Sie ein 1 mm 2 Box, die durch Kalibrierung die Bilder bei 530 Pixeln gleich 1 mm bestimmt. überprüfen Sie, ob das freigestellte Bild platziert die Spitze der Kapillare Schleifen an der Spitze des Bildes. </li><li> Glätten Sie das so alle künftigen Bildanpassungen werden gleichmäßig aufgebracht werden Bilddas Bild. Dazu klicken Sie auf zu tun: Prozess Registerkarte 2D-Filter, glätten, BG Intensität der &quot;hellen&quot; Objektbreite auf &quot;75&quot;, anzuwenden. </li><li> Erhöhen Sie den Kontrast des Bildes so Kapillaren maximal visualisiert. Um dies zu tun, klicken Sie auf: Registerkarte Anzeige einzustellen, heben Gegensatz zum 75. </li><li> Flecken entfernen Sie das Bild, um die Kanten der Kapillaren, indem Sie auf Prozess Registerkarte 2D-Filter, Flecken entfernen, Kernel-Größe 7 x 7 glätten, gelten. </li><li> Finalisieren Sie den Bildkontrast so Kapillaren sind schwarz und der Hintergrund ist weiß. Führen Sie diesen Schritt durch Anpassen des Histogramms auf die &quot;best fit&quot; -Modell. Hinweis: Bitte beachten Sie in Repräsentative Ergebnisse Ein Beispiel für das, was das verarbeitete Bild sollte wie die folgenden Schritte aussehen Abbildung 2. </li></ol> 2. Darstellende Capillary Counts / Quantifizieren Kapillardichte <ol><li> Auf jedem Bild, manuell ein Teil eines gut definierten Kapillare mit dem &quot;Zielobjekt&quot; featur wählene erkannt als Objekte, die vom Programm gezählt werden. Wählen Sie dann einen kleinen Teil der Hintergrund, mit dem &quot;Hintergrund&quot; -Funktion, als Hinweis auf Bereiche, die durch die Zählalgorithmus nicht berücksichtigt werden müssen. Hinweis: Die Kombination dieser Highlights bewirkt, dass alle Kapillaren unter Vernachlässigung des Hintergrundrauschens hervorgehoben. Die Quantifizierung (Zählen) Protokoll nutzt Computer-Funktionen, um Teile der Basis von Farbe und Morphologie Bild zu unterscheiden. </li><li> Verwenden Sie die Funktion count, um sofort zu zählen alle Kapillaren in das Bild mit dem bildgebenden Geräten. Die minimalen Durchmesser der gezählten Objekte auf 5 Pixel, um zu vermeiden Zählen Störgeräusche Kapillaren. </li><li> Für jeden einzelnen, zählen die durchschnittlich 3-4 Bilder, um eine zuverlässigere Beurteilung zu erhalten. Hinweis: Bitte beziehen Sie sich auf 3 in Repräsentative Ergebnisse Ein Beispiel für das, was das Bild sollte wie während des Verfahrens Zählen aussehen. </li></ol&gt; 3. Erstellen und Verwenden von Makros, um Bildverarbeitung Automatisieren Hinweis: Um Zeit zu sparen, können Makros, um eine spezifische Abfolge von Prozessen auf einem oder mehreren Bildern automatisch durchzuführen erstellt werden. Diese Sequenzen können, um die Bildänderungen schneller zu machen, angefertigt werden. Im Wesentlichen erinnern diese Makros, wie die Bilder verarbeitet, und führen Sie alle Schritte schnell und ohne Benutzereingaben. Darstellende zählt auf 12 Kapillar-Bilder nimmt diese Labor zwischen 20 bis 30 Minuten mit den Makros (2 bis 3 Minuten pro Bild), im Gegensatz zu etwa 8 min pro Bild ohne die Makros. Daher ist mit den Makros 3-5 mal effizienter als manuelle Verarbeitung jedes einzelnen Bildes. <ol><li> Um ein Makro zu erstellen, wählen Sie &quot;Makro aufzeichnen&quot; und auf einem Bild führen Sie die Schritte und Verfahren gewünscht wird, wie in Schritt 1 und 2 oben beschrieben. Benennen Sie den Makro basierend auf dem die Bildverarbeitungsschritte für die Zukunft durchgeführt wurden. Wenn using das Makro auf zukünftige Bilder, klicken Sie einfach auf &quot;run-Makro&quot;, und das Programm wird automatisch die aufgenommenen Verbesserungen an der Bild (n) gewünschte Anwendung. Hinweis: Diese Übung verwendet ein Makro, um alle bis auf einen der Schritte in Abschnitt 1 der Methoden in wenigen Sekunden durchzuführen. Der einzige Schritt, der Benutzereingaben erforderlich ist die Wahl, wo das Bild zu bearbeiten, Schritt 1.2. </li></ol>

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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. 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The authors have no conflicts of interest.

Nailfold Kapillaroskopie zeigt Versprechen als ein klinisch nützliches Werkzeug in der Zukunft für verschiedene Onkologie, Herz-Kreislauf und rheumatologische Erkrankung Anwendungen. Die Bildaufnahme-Prozess ist ziemlich konstant unter den Forschern, doch gibt es derzeit mehrere Methoden zur Bildverarbeitung und Analyse. Methoden umfassen derzeit computergestützte und manuelle Kapillare zählt. Manuelle Zählungen sind problematisch, da sie zeitaufwendig sind und unter den Benutzer Subjektivität und Müdigkeit. Stro...

Mikrovaskuläre Bildgebung ist ein schnell wachsendes Gebiet mit vielen potentiellen klinischen Anwendungen. 2 beispielsweise Onkologen verwenden microvessel Bildgebung, um das Ausmaß der Tumorangiogenese zu ermitteln, wodurch man wertvolle Informationen über den Zustand des Tumors und Einblicke in mögliche Behandlungsoptionen. 3 4 Allerdings nailfold Kapillaroskopie ist vielleicht die kosteneffiziente und breit anwendbare Form von mikrovaskulären Bildgebung. Die Forscher sind mit Hilfe von Video nailfold Kapillaroskopie zur Blutflussraten zu studieren und zu untersuchen Kapillare Morphologie. 5 6 Video- und Standbild nailfold Kapillaroskopie sind Zusätze für Diagnose und Behandlung von Raynaud-Phänomen und verschiedene Erkrankungen des Bindegewebes wie systemische Sklerose kümmern. 2 Nailfold Kapillaroskopie hat verschiedene potentielle kardiovaskuläre Anwendungen. Die aktuelle Forschung mit nailfold Kapillaroskopie schlägtdass Diabetes mellitus Typ 1 und Typ 2-Patienten weisen eine hohe Prävalenz von abnormalen Kapillare Morphologie, haben jedoch unverändert Kapillare Dichten im Vergleich zu nicht-diabetischen Individuen. 7-8 Kapillaroskopie wurde auch experimentell bei Hypertonie untersucht. Struktur Kapillare Verdünnung, die zu einem reduzierten Kapillardichte wurde bei hypertensiven Individuen im Vergleich zu nicht-Hypertonie-Patienten nachgewiesen. 9-10 Im Unterschied zu diesen älteren hypertensiven Patienten (Durchschnittsalter 40 und älter), die Strukturverdünnung aufweisen, hat die neuere Forschung gezeigt, dass jüngere hypertensiven Patienten (Durchschnittsalter unter 40 Jahre alt) sind funktional Verdünnung ohne strukturelle Verdünnung. 11. Dies deutet darauf hin, dass funktionelle Verdünnung vor auftritt und über die Zeit nicht, um strukturelle rarefaction Fortschritte. / Indapamid Interessanterweise hypertensiven Patienten mit bestimmten blutdrucksenkenden Medikamenten, wie Perindopril behandelten angezeigt Normal Kapillardichte und endothelialen Funktion nach der Behandlung, während die mit ACE (Angiotensin-Converting-Enzym-Inhibitoren) oder Diuretika behandelt aufrechterhalten trotz vergleichbarer Blutdruckregelung eine geringe Kapillardichte. 12 Dies legt nahe, dass einige Antihypertensiva kann Kapillardichte durch Umkehren der Kapillare normalisieren Verdünnung durch Bluthochdruck verursacht werden. Darüber hinaus haben andere Forscher gezeigt, dass eine Verringerung der Salzaufnahme führt zu einer Umkehrung der sowohl funktionelle und strukturelle Kapillare Verdünnung bei hypertensiven Individuen. 13 Trotz der verschiedenen möglichen klinischen Anwendungen dieser Technologie, gibt es wenig Standardisierung in der Technik für die Quantifizierung Kapillardichte Bilder. 2 Bisher haben die Forscher festgestellt, dass Kapillardichte Ergebnisse reproduzierbar sowohl aus intra-observer und interBeobachterPerspektive nur, wenn die genaue gleichen Bereich wird jedes Mal gezählt. 1,14 15 </sup&gt; Zu beachten ist, haben frühere Forscher weitgehend durchgeführt Kapillare zählt manuell mit dem bloßen Auge, 9 16 17 18, die eine langsame und subjektiver Prozess ist. Standardisierten, computergestützte Algorithmen zur Quantifizierung der Kapillare Bilder theoretisch eine effizientere und reproduzierbare Datenanalyse mit weniger Subjektivität, die Erleichterung der klinischen Anwendung von Kapillaroskopie. Einige Forscher haben tatsächlich verwendeten Computer-Programme, die Daten von nailfold kapillaroskopische Bilder zu quantifizieren. 1,6 19 20 jeweils nur eine Veröffentlichung auf dem Laufenden beschreibt jedoch Zuverlässigkeit einer komplexen Software-Programm zur Verfügung zum Quantifizieren Kapillaroskopie Daten, 1, und dieses Programm wird als kompliziert zuvor oben durch die Forderung festgestellt, um die genaue selben Sichtfeld zu zählen. Wir stellen hier ein einfacheres, zuverlässiges Protokoll zum Quantifizieren von Kapillaren unter Verwendung eines standardisierten Algorithmus, der ermöglichtDie Verwendung von mehreren Sichtfeldern. Die Verwendung von mehreren Sichtfeldern vereinfacht nicht nur das Verfahren, sondern ermöglicht auch die Beurteilung der normalen biologischen Variation Kapillare Zahl. Das Ziel dieser Studie ist es, eine reproduzierbare und effiziente computerbasierten Algorithmus, der die Kapillare Quantifizierungsprozess standardisiert beschreiben. Während diese Verfahren sind noch nicht vollständig automatisiert sie benötigen sehr wenig Benutzereingabe und rasche und zuverlässige Quantifizierung der Bilder.

<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

Kapillaroskopie ist eine nicht-invasive, effizient, relativ kostengünstig und leicht zu erlernen Methode zur direkten Visualisierung der Mikrozirkulation. Die Kapillaroskopie Technik kann einen Einblick in mikrovaskulärer die Gesundheit des Patienten, was zu einer Vielzahl von potentiell wertvollen dermatologischen, ophthalmologischen, rheumatologische und kardiovaskulären klinischen Anwendungen. Zusätzlich kann das Tumorwachstum angiogeneseabhängig, die durch Messung Gefäßdichte innerhalb der Tumor quantifiziert werden kann. Allerdings gibt es derzeit wenig bis keine Standardisierung von Techniken und nur eine Veröffentlichung Zeitpunkt meldet die Zuverlässigkeit eines derzeit verfügbaren komplexen Computer basierte Algorithmen zum Quantifizieren Kapillaroskopie Daten. 1 Dieser Artikel beschreibt ein neues, einfacheres, zuverlässige, standardisierte Kapillare Zählalgorithmus zur Quantifizierung nailfold Kapillaroskopie Daten. Eine einfache, reproduzierbare computerisierte Kapillaroskopie Algorithmus, wie dies mehr zu erleichternweit verbreiteten Einsatz der Technik bei den Forschern und Klinikern. Viele Forscher derzeit analysieren Kapillaroskopie Bilder von Hand, die Förderung der Ermüdung des Benutzers und Subjektivität der Ergebnisse. Dieser Aufsatz beschreibt eine neue, einfach zu bedienende automatischen Bildverarbeitungsalgorithmus, zusätzlich zu einem reproduzierbaren, halbautomatisierten Zählalgorithmus. Dieser Algorithmus ermöglicht die Analyse von Bildern in Minuten bei gleichzeitiger Reduzierung der Subjektivität; nur eine minimale Menge von Trainingszeit (nach unserer Erfahrung, weniger als 1 h) erforderlich, um die Technik zu erlernen.

Das Ziel dieses Bildverarbeitungsverfahrens ist es, die Kapillaren vom Hintergrundbild unterscheiden, so dass sie genau quantifiziert werden kann. Sowohl unvollständige Bildverarbeitung und Bildverarbeitung übermäßiger wirken sich nachteilig auf das Programm in der Lage ist die Kapillaren zu quantifizieren. Wie in Figur 3 zu sehen ist, macht unvollständige Bildverarbeitungs die Kapillaren schwierig aus dem Hintergrund zu unterscheiden. Entscheidend ist, dass der Benutzer in der Lage, leicht zu unte...

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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.

Reproduzierbarer computerisiertes Verfahren zur Quantifizierung der Kapillardichte Verwendung nailfold Kapillaroskopie

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 Aufrufe.  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. 

Serie: 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.

Photothrombotic Ischämie: eine minimal-invasive und reproduzierbare photochemische kortikale Läsion Modell für 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 ...

Forschung

Medizin

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.

Generation und 3-Dimensional Quantifizierung der arterielle Läsionen in Mäuse mit optischen Projektionstomographie

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.

Die Quantifizierung von Intraperitonealtest 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 ...

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.

Methode der direkten segmentale Intra-Leber-Lieferung Mit einer Rattenleber Hilar Clamp Modell

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.

Reproduzierbare Arterielle Denudation Verletzung von Infra Abdominal Aortic Spannen in einem Mausmodell

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.

Ein Verfahren zur Quantifizierung der oberen Extremität Leistung im täglichen Leben Accelerometer Verwendung

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 von ischämischen ventrikuläre Tachykardie mit einer multipolaren Katheter und 3-dimensionale Mapping-System für High-Density-Electro-anatomische Rekonstruktion

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.

Ein computergestütztes 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 ...

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.

Ein reproduzierbares intensivstationsorientiertes Endotoxinmodell bei Ratten

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