Diese Arbeit wurde von Grant Award Numbers R01HL096593 und K23HL72825 vom National Heart, Lung, and Blood Institute unterstützt. Der Inhalt ist ausschließlich der Verantwortung der Thomas Jefferson University und nicht notwendigerweise die offizielle Meinung des National Heart, Lung, and Blood Institute oder dem National Institutes of Health.

Fallbericht (erforderlich, falls zutreffend): NA, das ist immer noch eine experimentelle Forschung Verfahrens, noch nicht klinisch eingesetzt. Diagnose, Beurteilung und Plan (erforderlich, falls zutreffend): NA, das ist immer noch eine experimentelle Forschung Verfahrens, noch nicht klinisch eingesetzt. Procedure (erforderlich): Dieser Teil sollte eine Schritt-für-Schritt-Beschreibung der relevanten Verfahren gehören, erfüllen die Richtlinien unten. Ein. Kapillar-Mikroskopie (Abbildung 1) <ol><li> Unsere Kapillaroskopie Technik wurde von Serne und seine Kollegen 1 angepasst. Ein Ausschlusskriterium für dieses Verfahren ist Kollagenosen, da Kollagenosen produziert bekannt Kapillare Veränderungen 2. </li><li> Nach einer mindestens 10-hr nächtlichem Fasten und 20 min von sitzenden übrigen sind mikrovaskuläre Messungen für eine halbe Stunde zwischen 7 durchgeführtund 11 Uhr, in einer ruhigen, temperierten Raum (aufrechterhalten zwischen 21,5-22,5 ° C), mit dem Thema in der Sitzposition und die linke Hand auf Herzhöhe. </li><li> Nagelfalz Kapillaren in die Rückenhaut des dritten Fingers visualisiert mit einem Stereomikroskop (Olympus, Center Valley, PA), mit einem 4-Megapixel-SPOT Insight monochrome Digitalkamera (Modell-Nummer IN-1400: Diagnostic Instruments, Sterling Heights, MI) verbunden ist, und einen Laptop (Dell Latitude D600: Dell, Austin, TX). Um die Bewegung zu begrenzen, werden die linke Hand und Unterarm lose mit einem gefalteten Decke bedeckt, und ruhte auf einem anderen gefalteten Decke an der Basis des Mikroskops positioniert. </li><li> Nagelbett Beleuchtung mit einer 250-W-Halogenlampe Glasfaser Lampe (KL 2500LCD:; Elmsford, NY Schott-Fostec); erreicht zusätzliche Beleuchtung aus einer ergänzenden 150W LWL Halogen-Lichtquelle (B &amp; B Mikroskope, Ltd, Warrendale, PA) verwendet wird bei dunkelhäutigen Personen. </li><li> Um die Kapillaren, die Visualisierung3.2x Objektiv (Olympus 3.2/0.07) mit einem Gesamtsystem Vergrößerung 38.4x verwendet. </li><li> Mit SPOT Imaging-Software mit der Kamera, Licht / Dunkel-Kontrast in der Kapillare Fotografien verstärkt wird unter Verwendung der gleichen Standard-SPOT Software-Funktion (Streckung von hellen und dunklen Bereichen), um die Sichtbarkeit der Kapillaren in allen Fächern zu maximieren. </li><li> Um Kapillardichte quantifizieren, werden digitale Aufnahmen alle 3-5 sec in jeder der drei Stufen genommen, bei Ruhebasiswert während postokklusiven reaktive Hyperämie, und während Venenverschluss. (A) Bei Ruhebasiswert sind Aufnahmen über einen Zeitraum von drei Minuten zu erkennen Kapillaren in Ruhe durchströmt genommen. (B) Während postokklusiven reaktive Hyperämie, werden Aufnahmen gemacht, um funktionell perfundiert Kapillaren (baseline plus Reserve Kapillaren) zu quantifizieren, wie folgt. Zuerst wird eine Okklusion Manschette auf den linken Oberarm bis 40 mm Hg über systolische Druck für 10 min aufgepumpt. Mikrofotografien werden dann während th genommene ersten Minuten unmittelbar nach Freigabe der arteriellen Verschlusskrankheit, Visualisieren Alle funktional perfundiert Kapillaren. Lower Kapillardichte nach reaktive Hyperämie zeigt beeinträchtigt funktionelle Kapillardichte Rekrutierung und somit funktionale Verdünnung. (C) Während Venenverschluss Mikrophotographien getroffen werden, um eine maximale Kapillardichte, die sowohl perfundierten (mit aktiver roten Blutkörperchen (RBC) motion) und perfundierte (gefüllt mit stehendem, nicht bewegenden RBCs) Kapillaren 3 wie folgt enthält quantifizieren. Nach zehn Minuten Pause nach der reaktiven Hyperämie postokklusiven Verfahren wird der Armmanschette bis 60 mm Hg für 60 Sek. aufgeblasen, passiv dadurch Blut in alle Kapillaren vorliegenden Patentanmeldung und Mikroaufnahmen wurden während dieser Zeit genommen. Da maximalen Kapillardichte umfasst alle Kapillaren vorliegenden strukturell eine Reduzierung der maximalen Kapillardichte zeigt strukturelle Verdünnung. </li><li> Kapillardichte wird als die Anzahl von Kapillaren pro Quadrat definiert millimeter der Nagelfalz Haut, und wird als Mittelwert aus vier Messungen aus den vier klar fokussierte Bilder, mindestens durch Bewegung verzerrt erhalten berechnet. In unseren Studien haben typische Werte für die Kapillar-counts (Kapillaren / mm 2) waren 55-80 für Baseline, 65-90 für die post-ischämische und 90-105 für venöse Okklusion. Die Werte für Prozent Kapillare Rekrutierung liegen typischerweise zwischen 5% und 25% (Mittelwert ~ 10-15%) und für Prozent perfundiert Kapillaren zwischen 70% und 95% (Mittelwert ~ 80-90%), wobei die Werte niedriger bei Hypertonikern als Normotonikern. Die Reproduzierbarkeit der Zählvorgang mit drei Beobachter, die unabhängige manuelle Einschätzungen von Fotografien von 10 verschiedenen Themen (Abbildung 2) durchgeführt wurden überprüft. Die Beobachter waren auf die Identität und den Blutdruck dieser Themen geblendet. Nach seiner Ausbildung, zeigte Folgezählungen durchgeführt unabhängig voneinander ein hohes Maß an Übereinstimmung. Durchschnittliche Inter-Rater-und Intra-rater Diskrepanzen der order von 2-3 Kapillaren / mm 2, und Intraklassenkorrelation Koeffizienten waren alle größer als 0,90 ist. Kurzfristige Änderung der Kapillaroskopie Maßnahmen waren von der gleichen Größenordnung wie Inter-Rater-und Intra-Rater-Abweichungen (ca. 2 Kapillaren / mm 2), aber längerfristige Variation über 2-3 Jahre beobachtet wurde, war eine Größenordnung größer (ca. 15 Kapillaren / mm 2), die anzeigt, dass Längenänderungen leicht von rater Variation unterscheiden. Zuverlässigkeit der beiden Kapillarwirkung Maßnahmen war ebenfalls hoch (Intraclass Korrelationskoeffizient = 0,84 für Prozent Kapillare Rekrutierung und 0,82 für Prozent perfundiert Kapillaren). </li><li> Die Ermittler jetzt nutzen einen Computer-basierten Methode zur Quantifizierung Kapillardichte mit Image-Pro Plus Imaging-Software (Version 6.2, Media Cybernetics, Inc., Bethesda, MD: Abbildung 3). Pearson Korrelationen zwischen Baseline, post-ischämische und venouns Staus zählt mit der Software und den entsprechenden manuellen Zählungen in 10 Themen durchgeführt wurden 0,78, 0,78 und 0,71 jeweils (alle p &lt;0,05), was darauf hinweist vernünftige Einigung zwischen den beiden Methoden. Zuverlässigkeit der computerbasierten Zählungen ist geringfügig niedriger als die der manuellen Zählungen aber immer noch hoch (Intraklassen Korrelationskoeffizient = 0,91 für Grundlinie, 0,86 für post-ischämische, und 0,84 für venöse Okklusion). Wir haben unveröffentlichte Daten zeigt doch die Vereinigung von automatisierten zählt mit multiplen kardiovaskulären Risikofaktoren wie Bluthochdruck, die wir derzeit zur Veröffentlichung. </li><li> Tabelle 1 fasst die Kapillardichte Messungen und Berechnungen. Einstellungsverfahren Prozent Kapillare wird durch Teilen der Erhöhung Kapillardichte durch postokklusiven reaktive Hyperämie (postokklusiven reaktive Hyperämie Kapillardichte minus Basislinie Kapillardichte) induziert wird, durch die maximale Kapillardichte (beobachtet während des passiven veno beurteiltuns Okklusion). Prozent perfundiert Kapillaren für den Anteil aller Kapillaren Gegenwart, perfundiert (funktionell aktiv) werden, und durch Dividieren postokklusiven reaktive Hyperämie Kapillardichte durch die maximale Kapillardichte berechnet. Beide Prozent Kapillare Rekrutierung und Prozent perfundiert Kapillaren spiegeln die Anzahl der funktionellen Kapillaren. Niedrigere Werte für diese Maßnahmen zeigen, funktionelle Kapillardichte Verdünnung. </li></ol> 2. Endothelfunktion Beurteilung <ol><li> Endothelfunktion wird vor und nach postokklusiven reaktive Hyperämie zu beurteilen, mit nicht-invasiven Messungen von Plethysmographie Unterarm Blutfluss, die gemäß dem Verfahren von Sivertsson, 4, welche die Endothel-abhängige Stimulus der reaktiven Hyperämie um Vasodilatation zu induzieren nutzt. </li><li> Mit dem Subjekt in der Sitzposition nach 10 min der Rückenlage Rest, gestreckt eine quecksilberfreie in Gummi Dehnungsmesser bis 10% über ihre Ruhelänge aroun umschlungenend des Subjekts Unterarm 5 cm unter der Ellenbeuge. </li><li> , Die wiederum mit einem Doppler-Recorder (CW-1; DE Hokanson, Inc; Belleveue, WA) verbunden ist;: die Dehnungsmessstreifen auf einen Plethysmograph (Redmond, WA DE Hokanson, Inc EC-4) verbunden ist. </li><li> Ein Oberarm Okklusion Manschette angelegt ist, und der Arm ist komfortabel auf Herzhöhe suspendiert Verwendung einer Schlinge verbunden Bandage auf einen einstellbaren intravenöse Pol. Systolischen und diastolischen Blutdruck und Herzfrequenz werden mit einem Dinamap ProCare 100 automatische BP Manschette (GE Healthcare, Piscataway, NJ) auf der gegenüberliegenden Arm gelegt erhalten. </li><li> Eine pädiatrische Manschette um das Handgelenk auf 200 mm Hg zu fließen, um die Hand zu okkludieren aufgeblasen. Die Oberarmmanschette wird auf 50 mm Hg aufgepumpten entleerten für 1,5 sec, und dann wieder aufgeblasen rasch vor jedem Unterarm Blutflussmessung, durch Expansion des Dehnungsmeßstreifens um den Unterarm platziert erhalten. </li><li> Unterarm Blutfluss (FBF) bei ruhte Baseline (FBF Basis) gemessen und wieder eint postokklusiven Hyperämie-induzierte maximale Vasodilatation (FBF max). Für Baseline Blutfluss Messungen werden vier aufeinander FBF Kurven innerhalb von 30 Sekunden (FBF Basis) erhalten. </li><li> Die Okklusionsvorrichtung Manschette wird dann auf 40 mm Hg über systolische Druck für 10 min aufgepumpt. Nach Freisetzung von Arterienverschluss (postokklusiven reaktive Hyperämie) werden vier aufeinanderfolgende FBF Kurven innerhalb der ersten 30 sec Strömungsrichtung (FBF max) erhalten. </li><li> Das Verhältnis FBF max / FBF Base wird als eine Schätzung der Vasodilatation berechnet, indem der Mittelwert der vier FBF max-Werte durch den Mittelwert der vier Basiswerte FBF. 5 Unterarm Gefäßwiderstand bei maximaler Vasodilation (FVR max) als das berechnet wird mittleren arteriellen Druck (MAP) durch FBF max unterteilt. FBF max während der reaktiven Hyperämie wird direkt an FBF nach maximal Infusion von intra-arterielle Acetylcholin, einem Endothel-abhängige Vasodilatator verwandt. 6 </sup&gt; Dementsprechend FBF max und das Verhältnis FBF max / FBF Basis sind nicht-invasive Maßnahmen der endothelialen Funktion. 6-8 Darüber hinaus akzeptiert, spiegeln sowohl FBF max und FVR max Widerstand Arterie strukturellen Veränderungen (erhöhte Wand / Lumen-Verhältnis). 9 </li></ol>

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Die Autoren haben keine finanziellen Interessenkonflikte im Zusammenhang mit dem Verhalten oder Veröffentlichung dieser Arbeit offen zu legen.

Kapillaroskopie (Kapillarmikroskopie) ein Nicht-Standard-Maß Kapillarstruktur. Doch derzeit gibt es keine Standard-Methoden für die direkte Beurteilung der Kapillare structure.Furthermore hat Kapillarmikroskopie weithin für die direkte Auswertung der capillarydensity in einer großen und wachsenden Körper der veröffentlichten Arbeiten 12, 13, 10, 11, 14-18 verwendet . Darüber hinaus haben wir die Kapillare Mikroskopie-Technik durch Korrelation Kapillaroskopie Erkenntnisse mit Unterarm Durchblutung, ein ...

non-invasive assessment of microvascular and endothelial function

Die Autoren haben Kapillaroskopie und Unterarm Blutfluss Techniken verwendet werden, um die Rolle in der Pathogenese mikrovaskuläre Dysfunktion von kardiovaskulären Erkrankungen untersuchen. Kapillaroskopie ist eine nicht-invasive, relativ kostengünstige Methode zur direkten Visualisierung der Mikrozirkulation. Einstellungsverfahren Prozent Kapillare wird durch Teilen der Erhöhung Kapillardichte durch postokklusiven reaktive Hyperämie (postokklusiven reaktive Hyperämie Kapillardichte minus Basislinie Kapillardichte) induziert wird, durch die maximale Kapillardichte (beobachtet während des passiven Venenverschluss) beurteilt. Prozent perfundiert Kapillaren für den Anteil aller Kapillaren Gegenwart, perfundiert (funktionell aktiv) werden, und durch Dividieren postokklusiven reaktive Hyperämie Kapillardichte durch die maximale Kapillardichte berechnet. Beide Prozent Kapillare Rekrutierung und Prozent perfundiert Kapillaren spiegeln die Anzahl der funktionellen Kapillaren. Der Unterarm des Blutflusses (FBF) Technik provides akzeptiert nicht-invasive Maßnahmen der endothelialen Funktion: Das Verhältnis FBF max / FBF Base wird als eine Schätzung der Vasodilatation berechnet, indem der Mittelwert der vier FBF max-Werte durch den Mittelwert der vier Basiswerte FBF. Unterarm Gefäßwiderstand bei maximaler Vasodilatation (FVR max) als mittleren arteriellen Druck (MAP) durch FBF max dividiert. Sowohl die Kapillaroskopie und Unterarm-Techniken sind leicht für den Patienten akzeptabel und kann schnell erlernt werden. Die mikrovaskuläre und endotheliale Funktion misst unter Verwendung der Methoden in diesem Papier beschriebene zukünftige Nutzen in klinischen Patienten kardiovaskuläre Risiko-Reduktion Strategien haben. Wie wir veröffentlicht haben Berichte zeigen, dass mikrovaskuläre und endotheliale Dysfunktion in der Anfangsphase der Hypertonie einschließlich prehypertension gefunden werden kann und mikrovaskulären endothelialen Funktion Maßnahmen schließlich Früherkennung helfen, Risiko-Stratifizierung und Prävention von end-stage Gefäßpathologie mit ihren potenziell tödlichen Folgen.

Unterschiede im Aussehen der Mikrovaskulatur zwischen normotonen und hypertonen Individuen sind leicht ersichtlich durch Vergleichen den 4 und 5. Abbildung 4 zeigt den typischen Netzwerk von Kapillaren in gerader gut organisierte Zeilen in einem normotensiven Individuum. Im Gegensatz dazu zeigt Abbildung 5 eine ungeordnete Muster der geschrumpften, coiled Kapillaren. Die Autoren haben ein anhaltendes Interesse an der Rolle der mikrovaskulär...

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

Non-invasive Assessment of Microvascular and Endothelial Function

Kapillaroskopie ist eine nicht-invasive, relativ kostengünstige Methode zur direkten Visualisierung der Mikrozirkulation. Der Unterarm Blutfluss Technik bietet akzeptiert nicht-invasive Maßnahmen der endothelialen Funktion.

Non-invasive Beurteilung der mikrovaskulären und Endothelfunktion

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

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

Research

JoVE Journal

Medicine

16.1K Aufrufe.  Thomas Jefferson University. Kapillaroskopie ist eine nicht-invasive, relativ kostengünstige Methode zur direkten Visualisierung der Mikrozirkulation. Der Unterarm Blutfluss Technik bietet akzeptiert nicht-invasive Maßnahmen der endothelialen Funktion.

Serie: Non-invasive Assessment of Microvascular and Endothelial Function

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

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

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

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

Ein In vivo Nager-Modell der Kontraktion-induzierte Verletzung und nicht-invasive Überwachung von Recovery

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

Forschung

Medizin

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

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

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

Non-invasive optische Messung des zerebralen Metabolismus und Hämodynamik bei Säuglingen

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

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

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

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

Murine Spinotrapezius Modell, um die Auswirkungen der Arteriolen Ligation auf mikrovaskuläre Funktion und Remodeling beurteilen

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

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

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

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

Einen methodischen Ansatz, um Nicht-invasive Assessments der Gefäßfunktion und Morphologie

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

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

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

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

Nicht-invasive Beurteilung der Wirksamkeit neuer Therapeutika für Darmpathologien Verwenden der seriellen endoskopische Bildgebung von lebenden Mäusen

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

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

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

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

Ultraschall Beurteilung der Endothelfunktion: A Technische Richtlinie der Fluss-vermittelten Dilatation-Test

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

Non-invasive Assessments of Subjective and Objective Recovery Characteristics Following an Exhaustive Jump Protocol

Fast recovery after strenuous exercise is important in sports and is often studied via cryotherapy applications. Cryotherapy has a significant vasoconstrictive effect, which seems to be the leading factor in its effectiveness. The resulting enhanced recovery can be measured by using both objective and subjective parameters. Two commonly measured subjective characteristics of recovery are delayed-onset muscle soreness (DOMS) and ratings of perceived exertion (RPE). Two important objective recovery characteristics are countermovement jump (CMJ) performance and peak power output (PPO). Here, we provide a detailed protocol to induce muscular exhaustion of the frontal thighs with a self-paced, 3 x 30 countermovement jump protocol (30-s rest between each set). This randomized controlled trial protocol explains how to perform local cryotherapy cuff application (+ 8 &#176;C for 20 min) and thermoneutral cuff application (+ 32 &#176;C for 20 min) on both thighs as two possible post-exercise recovery modalities. Finally, we provide a non-invasive protocol to measure the effects of these two recovery modalities on subjective (i.e., DOMS of both frontal thighs and RPE) and objective recovery (i.e., CMJ and PPO) characteristics 24, 48, and 72 h post-application. The advantage of this method is that it provides a tool for researchers or coaches to induce muscular exhaustion, without using any expensive devices; to implement local cooling strategies; and to measure both subjective and objective recovery, without using invasive methods. Limitations of this protocol are that the 30 s rest period between sets is very short, and the cardiovascular demand is very high. Future studies may find the assessment of maximum voluntary contractions to be a more sensitive assessment of muscular exhaustion compared to CMJs.

This protocol describes the procedure for non-invasive recovery assessments during a 72 h recovery period. This protocol induces muscular exhaustion of the frontal thighs using countermovement jumps and uses either local cold-cuff or thermoneutral-cuff application as a recovery modality.

Nicht-invasive Assessments von subjektiven und objektiven Erholungsmerkmalen nach einem erschöpfenden Sprungprotokoll

Fast recovery after strenuous exercise is important in sports and is often studied via cryotherapy applications. Cryotherapy has a significant ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

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

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

Bewertung der menschlichen Fettgewebe mikrovaskuläre Funktion mit Videomicroscopy

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

Non-invasive Assessment of Dorsiflexor Muscle Function in Mice

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

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

Nicht-invasive Beurteilung des releveurs Muskelfunktion bei Mäusen

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

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

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

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