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.

Nota: Il processo di acquisizione per ottenere immagini capillari è stato pubblicato in precedenza ed è realizzato utilizzando una fotocamera digitale con un programma informatico di acquisizione e analisi delle immagini corrispondente 11 21 Questo laboratorio utilizza immagini fisse per l&#39;analisi, non video, semplificando l&#39;acquisizione di immagini per l&#39;analisi.. Quanto segue descrive la nuova tecnica per quantificazione dei capillari dalle immagini. 1. Miglioramento immagine di processo <ol><li> Ottenere immagini digitali con una macchina fotografica digitale in bianco e nero. Calibrare le immagini a un oggetto di dimensioni note per scattare una foto di un oggetto di lunghezza nota, come un righello con la fotocamera. Questo processo permette il programma informatico per misurare e contare capillari accuratamente dopo l&#39;elaborazione. L&#39;ideale per la massima precisione, dovrebbe essere utilizzato un reticolo (brano scientificamente fabbricato di vetro con un righello inciso in esso). Misurare il numero di pixel in una piazza 1 mmbox utilizzando un programma per computer. Nota: La chiave per la riproducibilità e la standardizzazione di questo protocollo dipende in gran parte il corretto posizionamento della scatola 1 mm 2 che viene conteggiato. </li><li> Utilizzare strumenti di miglioramento di contrasto per scurire i capillari e schiarire lo sfondo, che ci permetterà di ottimizzare la visualizzazione dei capillari. Differenziazione iniziale dei capillari di sfondo è importante per il corretto ritaglio dell&#39;immagine nei passaggi successivi. A tale scopo, regolando l&#39;immagine con un Per fare ciò cliccate su &quot;miglior istogramma in forma.&quot;: La cattura, l&#39;intensità, l&#39;immagine istogramma, meglio si adattano. </li><li> Ritagliare una regione di interesse (ROI) per il conteggio capillare come nuova immagine (selezionate, regioni di interesse). Utilizzare una scatola 1 mm 2, che è stata determinata calibrando le immagini a 530 pixel eguagliando 1 mm. verificare che l&#39;immagine ritagliata pone l&#39;apice delle anse capillari in cima dell&#39;immagine. </li><li> Appiattire l&#39;immagine in modo che tutti i futuri regolazioni dell&#39;immagine saranno applicate in modo uniformel&#39;immagine. Per fare ciò cliccate su: scheda processo, filtri 2D, appiattire, intensità BG di &quot;brillante&quot;, object width a &quot;75,&quot; si applicano. </li><li> Aumentare il contrasto dell&#39;immagine in modo capillari vengono visualizzate al massimo. Per fare ciò cliccate su: scheda, visualizzazione regolare, aumentare il contrasto a 75. </li><li> Despeckle l&#39;immagine per smussare i bordi dei capillari cliccando su scheda processo, filtri 2D, smacchiatura, la dimensione del kernel 7 x 7, si applicano. </li><li> Finalizzare il contrasto dell&#39;immagine in modo capillari sono neri e lo sfondo è bianco. Eseguire questo passaggio regolando l&#39;istogramma per il modello &quot;best fit&quot;. Nota: Si prega di fare riferimento alla figura 2 a Rappresentante dei risultati per un esempio di ciò che l&#39;immagine elaborata dovrebbe essere simile seguendo questi passaggi. </li></ol> 2. Esecuzione Conti capillari / quantificazione capillare Densità <ol><li> Su ogni immagine, selezionare manualmente una parte di un capillare ben definito utilizzando il featur &quot;target oggetto&quot;e di essere riconosciuti come oggetti da contare dal programma. Quindi, selezionare una piccola parte del fondo, utilizzando la funzione &quot;background&quot;, come riferimento per aree che devono essere prese in considerazione dall&#39;algoritmo conteggio. Nota: La combinazione di questi punti salienti fa sì che tutti i capillari da evidenziare, trascurando il rumore di fondo. Il protocollo di quantificazione (conteggio) utilizza le funzioni del computer di differenziare le parti dell&#39;immagine in base al colore e morfologia. </li><li> Utilizzare la funzione di conteggio per contare istantaneamente tutti i capillari nell&#39;immagine con il apparecchiature di imaging. Impostare il diametro minimo di oggetti contati a 5 pixel in modo da evitare di contare il rumore di fondo come capillari. </li><li> Per ogni individuo, contare il media di 3 - 4 immagini in modo da ottenere una valutazione più affidabile. Nota: Si prega di fare riferimento alla Figura 3 a Rappresentante dei risultati per un esempio di ciò che l&#39;immagine dovrebbe essere simile nel corso della procedura di conteggio. </li></ol&gt; 3. Creazione e utilizzo di macro per automatizzare Image Processing Nota: Per risparmiare tempo, macro possono essere create per eseguire automaticamente una sequenza specifica di processi su uno o più immagini. Queste sequenze possono essere personalizzati al fine di apportare le modifiche di immagine più veloce. In sostanza, queste macro ricordano come le immagini vengono elaborate, ed eseguire tutti i passaggi in modo rapido e senza l&#39;input dell&#39;utente. Esecuzione conta su 12 immagini capillari prende questo laboratorio tra i 20 ei 30 minuti con le macro (da 2 a 3 min per immagine), a differenza di circa 8 minuti per foto senza le macro. Pertanto, utilizzando le macro è da 3 a 5 volte più efficiente di elaborazione manualmente ogni singola immagine. <ol><li> Per creare una macro, selezionare &quot;macro record&quot; e un&#39;immagine eseguire le fasi ei processi desiderati, come descritto ai punti 1 e 2 di cui sopra. Nome della macro in base alla quale l&#39;elaborazione delle immagini fasi sono state eseguite per riferimento futuro. Quando using la macro sulle immagini future, è sufficiente cliccare su &quot;macro run&quot; e il programma applicherà automaticamente i miglioramenti registrati all&#39;immagine desiderato (s). Nota: Questo laboratorio utilizza una macro per eseguire tutti, ma uno dei passi nella Sezione 1 dei metodi in pochi secondi. L&#39;unico passo che richiede l&#39;input dell&#39;utente è scegliere dove tagliare l&#39;immagine, Step 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. 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The authors have no conflicts of interest.

Capillaroscopia ungueale mostra la promessa come uno strumento clinicamente utile in futuro per vari oncologia, cardiovascolare, e le applicazioni di malattie reumatologiche. Il processo di acquisizione delle immagini è abbastanza coerente tra i ricercatori, ma attualmente ci sono diversi metodi per l&#39;elaborazione delle immagini e di analisi. I metodi attualmente includono i conteggi capillari informatiche e manuali. Conteggi manuali sono problematici in quanto sono in termini di tempo, e soggetto alla soggettivit?...

Imaging microvascolare è un settore in rapida crescita, con molte potenziali applicazioni cliniche. 2, ad esempio, oncologi utilizzano immagini dei microvasi per determinare l&#39;entità di angiogenesi tumorale, ottenendo preziose informazioni sullo stato del tumore e comprensione possibili opzioni di trattamento. 3 4 Tuttavia, ungueale capillaroscopia è forse la forma più efficiente e ampiamente applicabile costo dell&#39;imaging microvascolare. I ricercatori stanno utilizzando il video letto ungueale capillaroscopia per studiare i tassi di flusso di sangue e indagare la morfologia capillare. 5 6 Sia video e immagini fisse ungueale capillaroscopia sono aggiunte per la cura per diagnosticare e trattare fenomeno e di varie malattie del tessuto connettivo di Raynaud come la sclerosi sistemica. 2 Capillaroscopia ungueale ha varie potenziali applicazioni cardiovascolari pure. La ricerca attuale suggerisce usando letto ungueale capillaroscopiache diabete mellito di tipo 1 e di tipo 2 pazienti mostrano una elevata prevalenza di anomalie morfologiche dei capillari, ma hanno densità capillare invariati rispetto agli individui non diabetici. 7-8 capillaroscopia è stato studiato anche sperimentalmente in ipertensione. Rarefazione capillare strutturale portando ad una densità capillare ridotta è stata dimostrata in soggetti ipertesi, rispetto ai soggetti non ipertesi. 9-10 In contrasto con questi pazienti ipertesi anziani (età media 40 anni e sopra), che presentano rarefazione strutturale, la ricerca più recente ha dimostrato che pazienti ipertesi più giovani (età media sotto i 40 anni) hanno rarefazione funzionale senza rarefazione strutturale. 11 Ciò suggerisce che rarefazione funzionale si verifica prima e può evolvere nel tempo per la rarefazione strutturale. È interessante notare che i pazienti ipertesi trattati con specifici farmaci antipertensivi, come perindopril / indapamide visualizzata normaAl densità capillare e la funzione endoteliale dopo il trattamento, mentre quelli trattati con ACE (enzima di conversione-enzima) inibitori o diuretici mantenuto una bassa densità capillare, nonostante il controllo della pressione sanguigna comparabili. 12 Questo suggerisce che alcuni farmaci antipertensivi possono normalizzare la densità capillare invertendo la capillare rarefazione causata da ipertensione. Inoltre, altri ricercatori hanno dimostrato che una riduzione dell&#39;assunzione di sale porta ad inversione di funzionale e strutturale rarefazione capillare nei soggetti ipertesi. 13 Nonostante le varie potenziali applicazioni cliniche di questa tecnologia, c&#39;è poco standardizzazione tecnica per la quantificazione immagini densità capillare. 2 Ad oggi, i ricercatori hanno scoperto che i risultati di densità capillare sono riproducibili sia da un intra-osservatore e inter-osservatore prospettiva solo se l&#39;esatto stessa area viene contato ogni volta. 1,14 15 </sup&gt; Da segnalare, i ricercatori precedenti hanno in gran parte effettuato conta capillari manualmente utilizzando occhio nudo, 9 16 17 18, che è un processo lento e soggettiva. Standardizzati, basati su computer algoritmi per la quantificazione di immagini capillari teoricamente fornire analisi dei dati più efficiente e riproducibile con meno soggettività, facilitando applicazioni cliniche di capillaroscopia. Alcuni ricercatori hanno infatti utilizzato programmi computerizzati per quantificare i dati ungueale immagini capillaroscopiche. 1,6 19 20 Tuttavia, solo una pubblicazione ad oggi descrive l&#39;affidabilità di un programma di software complesso disponibile per quantificazione dati capillaroscopia, 1 e questo programma è complicato come precedentemente notato sopra dalla necessità di contare il campo visivo stessa identica. Qui, presentiamo un protocollo affidabile semplice per quantificazione capillari utilizzando un algoritmo standardizzato che consentel&#39;uso di più campi visivi. L&#39;uso di più campi visivi non solo semplifica la procedura, ma permette anche la valutazione della normale variazione biologica conteggio capillare. Lo scopo di questo studio è quello di descrivere un algoritmo computerizzato riproducibili ed efficiente che standardizza il processo di quantificazione capillare. Anche se questi metodi non sono completamente automatizzate richiedono molto poco l&#39;input dell&#39;utente, e forniscono quantificazione rapida e sicura delle immagini.

<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

Capillaroscopia è una metodologia non invasiva, efficiente, relativamente economico e facile da imparare per visualizzare direttamente la microcircolazione. La tecnica capillaroscopia può fornire indicazioni in salute microvascolare di un paziente, che porta a una varietà di dermatologica potenzialmente prezioso, oftalmologico, applicazioni cliniche reumatologiche e cardiovascolari. Inoltre, la crescita del tumore può dipendere angiogenesi, che può essere quantificata misurando la densità dei microvasi all&#39;interno del tumore. Tuttavia, vi è attualmente poco o nessun standardizzazione delle tecniche, e solo una pubblicazione aggiornata riporta l&#39;affidabilità di un attualmente disponibili algoritmi computerizzati, complessi di quantificazione dei dati capillaroscopia. 1 Questo articolo descrive un nuovo algoritmo, semplice, affidabile, capillare standardizzata conteggio per quantificazione dei dati capillaroscopia ungueale. Un algoritmo semplice, riproducibile computerizzato capillaroscopia come questo faciliterebbe moreuso diffuso della tecnica tra ricercatori e clinici. Molti ricercatori attualmente analizzano le immagini capillaroscopia a mano, promuovendo l&#39;affaticamento utente e la soggettività dei risultati. Questo documento descrive un romanzo, facile da usare automatizzato algoritmo di elaborazione delle immagini in aggiunta a un algoritmo di conteggio riproducibile, semi-automatico. Questo algoritmo consente di analisi di immagini in pochi minuti riducendo soggettività; solo una minima quantità di tempo di formazione (secondo la nostra esperienza, meno di 1 ora) è necessario per imparare la tecnica.

L&#39;obiettivo di questa procedura di elaborazione delle immagini è quello di differenziare i capillari dall&#39;immagine di sfondo in modo che possano essere quantificati con precisione. Entrambi elaborazione delle immagini incompleta e l&#39;elaborazione di immagini eccessive sono dannose per la capacità del programma di quantificare i capillari. Come si vede nella figura 3, l&#39;elaborazione delle immagini incompleta rende i capillari difficile distinguere dallo sfondo. E &#39;fondamentale che l&...

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.

Un metodo computerizzato riproducibile per la quantificazione della densità capillare con letto ungueale Capillaroscopia

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 Views.  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. 

Video: 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.

Ischemia Photothrombotic: un modello fotochimico lesione corticale minimamente invasiva e riproducibile per mouse Studi Stroke

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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Medicina

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.

Generazione e quantificazione 3-Dimensional di arteriosa lesioni nei topi mediante tomografia ottica di proiezione

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.

La quantificazione di intra-peritoneale del cancro ovarico metastasi

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.

Metodo di consegna diretta segmentale intra-epatico utilizzando un modello di fegato di ratto Hilar morsetto

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.

Riproducibile arteriosa Denudation Lesioni da infrarenale aortico addominale di serraggio in un modello murino

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.

Un metodo per la quantificazione degli arti superiori performance nella vita quotidiana che utilizzano accelerometri

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.

Ablazione della tachicardia ventricolare ischemica utilizzando un catetere Multipolar e sistema di mappatura 3-dimensionale per ricostruzione Electro-anatomica ad alta densità

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 programma computerizzato di valutazione e formazione delle competenze funzionali, destinato alle competenze funzionali quotidiane basate sulla tecnologia

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 modello di endotossina orientato all'unità di terapia intensiva riproducibile nei ratti

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