Name: evaluation of blood lactate and plasma insulin during high-intensity exercise by antecubital vein catheterization
Uploaded: 2018-05-18T12:30:00.000+00:00
Duration: 4 min 28 s
Description: Watch this Scientific Journal Video about Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization at JoVE.com

Die Autoren möchten Dr. P. Karagiannis für die Zeitung lesen und Professor T. Hoyo für seine technische Unterstützung zu erkennen.

Alle hier beschriebene Methoden wurden von der lokalen Ethikkommission genehmigt (Doshisha Universität Ethikkommission: 15033), und waren in strikter Übereinstimmung mit den Standards der Deklaration von Helsinki. Informierte Zustimmung wurde von allen Teilnehmern erhalten. Teilnehmer waren männliche Freizeitsportler (Alter, 20.1 ± 1,2 Jahre; Gewicht, 69,7 ± 6,2 kg; Höhe, 176.1 ± 5,8 cm; Körper Fett 13,6 ± 1,5 % und VO2max, 52.6 ± 6,6 mL/min1schnitzelfleisch1).
1. Aufbau des Protokolls Übung
Hinweis: Die vorliegende Studie bestand aus der Bewertung von Blut hormonelle und Metaboliten Dynamik durch Blutentnahme aus der Vene Antecubital während ein HIIE Protokoll.
<ol><li>Fragen Sie das Thema, eine warme, mit 5 min des Radsports in der Ergometer bei niedriger Intensität durchzuführen. Lassen Sie nach dem Warm-up das Thema für 5 min ruhen.</li>
	<li>Bitte das Thema der folgenden HIIE in das Bein Fahrradergometer durchzuführen: drei Anfälle von 30 s hoch-intensives Training, getrennt durch 4 min. Rest (Abbildung 1). Hinweis: Die 30 s-Kämpfe bei gleicher Intensität zu 90 % den macht-Durchschnitt von einer zuvor durchgeführten Wingate-Test festzulegen. Finden Sie an anderer Stelle8 für eine vollständige Beschreibung der Wingate-Test.</li>
	<li>Um die Arbeitsbelastung für die 30 s-Kämpfe zu berechnen, verwenden Sie die folgenden Gleichungen: Arbeit = Kraft x Abstand (Gleichung 1) Kraft = Masse x Beschleunigung (Gleichung 2) Arbeit = (Masse x Beschleunigung) x Abstand (Gleichung 3) Hinweis: In diesem Modell ist die Masse der Arbeitslast (kg), und die Beschleunigung beträgt 9,81 m/s2. Die Entfernung beträgt der Durchmesser der Räder (1,622 m). Da das Rad 3,7 zeitgesteuerte /peal Drehung dreht, ist der Abstand pro Pedal Umdrehung 3,7 x 1.662. Andererseits, weil die Trittfrequenz 90 u/min-1ist, haben wir ein Pedalabstand mit 90 zu multiplizieren. Arbeit = Arbeitslast x 9,81 x 3,7 x 1.622 x 90 (Gleichung 4) Belastung = Arbeit / (9,81 x 3,7 x 1.622 x 90) (Gleichung 5)</li>
</ol>(2) Katheterisierung
Hinweis: Rohre und vacutainer sollten im Vorfeld zu vermeiden, dass Sie Zeit verlieren beschriftet werden.
<ol><li>Wickeln Sie einen Druckverband am Arm für die Katheterisierung ausgewählt. Um dies zu tun, legen Sie eine elastisches Band auf der Rückseite der gewählten Arm. Dehnen die Band, machen Sie eine Schleife mit der Band um den Arm und binden Sie die elastischen.</li>
	<li>Wählen Sie die sichtbaren Antecubital Vene und Sterilisieren Sie die Haut mit Alkohol oder Ethanol.</li>
	<li>Führen Sie die Spitze der sterilisierte Kanüle in die Vene in einem ungefähren Winkel von 15° mit der Haut. Hinweis: Die Kanüle sollte von der proximalen, distalen Richtung eingefügt werden. Wenn Kanülierung erfolgreich ist, sollte eine retrograde Entnahme von Blut beobachtet werden.</li>
	<li>Sichern Sie die Kanüle mit der Hand, und entfernen Sie die Nadel sanft. Hinweis: Um den Winkel der Nadel während der Übung zu halten, können Baumwolle Gewebe zwischen Haut und Nadel verwendet werden. Befestigen Sie diese Struktur mit einem Band.</li>
	<li>Verbinden Sie ein Vacutainer mit Heparin und aspirat für die erste Blutprobe (ca. 2,5 mL). Unmittelbar nach kühlen Sie der Blutentnahme indem man es in eine Eisbox für nicht länger als 40 Minuten. Bitten Sie ein Assistent um Hilfe, wenn Blutgaswerte analysiert werden sollen.</li>
	<li>Schließen Sie einen Schlauch an den Katheter und Spritzen Sie Heparin auf die volle Länge des Rohres um eine Blutgerinnung zu vermeiden. Schließen Sie den Schmetterling.</li>
	<li>Mit Klebeband, stabilisieren Sie den Katheter. Stellen Sie sicher, dass an dieser Stelle die Arm-Beweglichkeit des Themas mit der Blutentnahme nicht stört. Um diese Bedingung zu bestätigen, werden Sie sicher, dass die Nadelposition jede Bewegung nicht betroffen ist.</li>
</ol>3. Blut Samplings
<ol><li>Entfernen Sie das Heparin mithilfe einer Vacutainer bis es nur Blut in den Katheter gibt. Wenn Blut Rückblende beobachtet wird, sammeln Sie die Probe wie in Schritt 2.5 beschrieben.</li>
	<li>Spritzen Sie nach der Probenentnahme Heparin wieder in den Katheter Schlauch Blutgerinnung zu vermeiden. Nach dieser Aktion kann Schritt 2.5 für mehrere Proben Sammlung wiederholt werden.</li>
	<li>Nach die letzte Probe erhoben werden, sorgfältig die Kanülierung Rohr zu entfernen und die Haut mit Alkohol reinigen. Um mögliche Blutungen zu vermeiden, bitten Sie, das Thema weiterhin sterilisiert Baumwolle an der Stelle wo die Katheterisierung gemacht wurde.</li>
	<li>Nachdem Blut in die Vacutainer-Röhrchen gesammelt, sorgfältig etwa 1,5 mL Blut zu einem 1,5 mL Schlauch und halten Sie die Rohre auf dem Eis.</li>
	<li>Maßnahme-Blut-Laktat aus dem restlichen Blut mit einem Laktat-Detektor. Am selben Tag erhalten Sie dem Plasma der Blutproben (rund 1,5 mL Blut).</li>
	<li>Für Plasma-Teilung Zentrifugieren der Blutproben für 5 min bei 5000 X g bei 4 oC, und Trennen des Überstands in ein neues 1,5 mL-Tube.</li>
	<li>Wiederholen Sie die Zentrifugation noch einmal um das Plasma zu reinigen.</li>
	<li>Nach der Trennung Plasma speichern Sie die Proben bei-80 oC bis die Hormon-Analyse.</li>
	<li>Das Plasma-Insulin mit einem ELISA-Kit wie pro des Herstellers Protokoll (siehe Tabelle der Materialien) zu messen.</li>
</ol>

<ol>
	<li>Wahl, P., Mathes, S., K&#246;hler, K., Achtzehn, S., Bloch, W., Mester, J. <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+active+vs.+passive+recovery+during+Wingate-based+training+on+the+acute+hormonal,+metabolic+and+psychological+response.">Effects of active vs. passive recovery during Wingate-based training on the acute hormonal, metabolic and psychological response.</a> Growth Horm IGF Res. 23 (6), 201-208 (2013).</li><li>Wahl, P., Mathes, S., Achtzehn, S., Bloch, W., Mester, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Active+vs.+passive+recovery+during+high-intensity+training+influences+hormonal+response.">Active vs. passive recovery during high-intensity training influences hormonal response.</a> Int J Sports Med. 35 (7), 583-589 (2014).</li><li>Nalbandian, H. M., Radak, Z., Takeda, M. <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+active+recovery+during+interval+training+on+plasma+catecholamines+and+insulin.">Effects of active recovery during interval training on plasma catecholamines and insulin.</a> J Sports Med Phys Fitness. , (2017).</li><li>Stolen, T., Chamari, K., Castagna, C., Wislff, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiology+of+Soccer.">Physiology of Soccer.</a> Sport Med. 35 (6), 501-536 (2005).</li><li>Bangsbo, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+physiology+of+soccer--with+special+reference+to+intense+intermittent+exercise.">The physiology of soccer--with special reference to intense intermittent exercise.</a> Acta Physiol Scand Suppl. 619, 1-155 (1994).</li><li>MacInnis, M. J., Gibala, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Physiological+adaptations+to+interval+training+and+the+role+of+exercise+intensity.">Physiological adaptations to interval training and the role of exercise intensity.</a> J Physiol. 595 (9), 2915-2930 (2017).</li><li>Emberts, T., Porcari, J., Dobers-Tein, S., Steffen, J., Foster, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Exercise+intensity+and+energy+expenditure+of+a+tabata+workout.">Exercise intensity and energy expenditure of a tabata workout.</a> J Sports Sci Med. 12 (3), 612-613 (2013).</li><li>Smith, J. C., Hill, D. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+energy+systems+during+a+Wingate+power+test.">Contribution of energy systems during a Wingate power test.</a> Br J Sports Med. 25 (4), 196-199 (1991).</li><li>Dotan, R., Bar-Or, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Load+optimization+for+the+Wingate+Anaerobic+Test.">Load optimization for the Wingate Anaerobic Test.</a> Eur J Appl Physiol Occup Physiol. 51 (3), 409-417 (1983).</li><li>Carrasco-Benso, M. P., 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=Human+adipose+tissue+expresses+intrinsic+circadian+rhythm+in+insulin+sensitivity.">Human adipose tissue expresses intrinsic circadian rhythm in insulin sensitivity.</a> FASEB J. 30 (9), 3117-3123 (2016).</li><li>Hatfield, D. L., Nicoll, J. X., Kraemer, W. J. <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+Circadian+Rhythm+on+Power,+Force,+and+Hormonal+Response+in+Young+Men.">Effects of Circadian Rhythm on Power, Force, and Hormonal Response in Young Men.</a> J Strength Cond Res. 30 (3), 725-732 (2016).</li></ol>

Die Autoren haben nichts preisgeben.

In dieser Studie die Antecubital Vene war katheterisiert und Blutproben wurden erfolgreich gesammelt, während ein hohes Intervall-Trainings. Als ein Beispiel für die Verwendung dieser Methodik wurden Blutproben für Plasma-Insulin für Blut-Laktat und (nach Plasma Trennung) analysiert. Der Hauptvorteil dieser Methode ist, dass es die Sammlung von Blutproben in kurzen Zeiträume ermöglicht. Zur gleichen Zeit hat es einige Einschränkungen. Zum Beispiel die begrenzte Übung Modalitäten, die seine Verwendung; die Anzahl der Assistent benötigt und geschultes Fachpersonal erforderlich; und das Risiko einer geringen Probenvolumen infolge niedriger Blutdruck in den Unterarmen.
Für eine erfolgreiche Blutentnahme Probe sollte das Thema Arm in statischer Position sein. Weil die Unterarm-Mobilität eingeschränkt ist, kann in seiner jetzigen Form die Methode nur auf ein paar Übungen Modalitäten angewendet. Wir beschrieben hier, wie man die Methode anwenden, wenn die Übung wird in einem Fahrradergometer durchgeführt, aber mit Änderung könnte es auf einem Laufband Laufband verwendet werden. In beiden Fällen braucht man Personal mit Erfahrung in der Blutentnahme.
Bezüglich der Übung Protokoll gibt es einige Überlegungen, die angesprochen werden sollten. Wenn mehrere Studien ausgewertet (z.B., mit und ohne aktive Erholung zwischen den Anfällen) sein wollen, ist der Moment der Entscheidung für die Prüfung entscheidend. Um tagaktive Hormonschwankungen10,11zu vermeiden, empfiehlt es sich, alle Versuche zur selben Stunde an verschiedenen Tagen durchzuführen. Darüber hinaus ist die Tageszeit und die Mahlzeiten verzehrt Stoffwechsel betroffen. Daher empfiehlt das Fasten vor den Tests. Darüber hinaus empfiehlt sich zur Verbesserung der Reproduzierbarkeit einer Eingewöhnungszeit mit der Übung Protokoll von den Probanden vor der Prüfung. In diesem Zeitraum sollten Versuchspersonen (schrittweise) ähnliche Übung zu den wichtigsten Test durchführen.
Zusammenfassend beschrieben wir eine effektive Methode um mehrere venöses Blutproben während des Trainings zu erhalten. Diese Methode ist nützlich für das Sammeln von Proben, die verwendet werden können, um mehrere Moleküle während des Trainings (z.B., Metaboliten, Hormone, Proteine und RNA) zu studieren.

Die Messung der endokrinen und metabolischen Marker während der körperlichen Aktivität ist von Bedeutung für das Verständnis der physiologischen Auswirkungen der verschiedenen Übung Modalitäten. Während einige Übung Modalitäten (z. B. hoher Intensität Intervall Übung (HIIE)) schwanken Blut-Metaboliten und Hormonspiegel in relativ kurzer Zeit1,2,3.
Viele Sportarten (z.B. Fußball, Basketball und Rugby) bestehen aus kurzfristig hoch-intensives Training durch niedrig intensivem Training und/oder passive Perioden4,5getrennt sind. HIIE zeigt Ähnlichkeiten mit den oben genannten Sportarten und ist daher weithin als eine Trainingsmethode für Athleten6verwendet. Darüber hinaus wegen seiner Ähnlichkeit mit Sport, wurde HIIE als Modell zur Übung Physiologie7zu studieren. Blutabnahme ist entscheidend für die physiologische Variablen zu messen, und es wird häufig verwendet für Forschungszwecke und die Beurteilung der körperlichen Verfassung. Blutabnahme während HIIE kann jedoch aufgrund der kurzen Zeitspanne zwischen Musterkollektionen technisch schwierig sein. Als Lösung für dieses Problem präsentieren hier wir eine Methode, um Blutproben von venösen Katheterisierung Antecubital kurz erhalten Zeiträume während Themen HIIE durchführen.

<table><tbody><tr><td>Cycle ergometer</td><td>Monark</td><td>MONARK Ergomedic 874 E</td><td></td></tr><tr><td>Catheter</td><td>Terumo</td><td>SR-FF2032</td><td></td></tr><tr><td>Vacutainer</td><td>Terumo</td><td>PZ-D03</td><td></td></tr><tr><td>Tube for catheter</td><td>Terumo</td><td>SP-PTW30L02</td><td></td></tr><tr><td>Heparin</td><td>Terumo</td><td>PF-10HF10UA</td><td></td></tr><tr><td>Insulin ELISA kit</td><td>Abnova</td><td>ELISA Kit KA0921</td><td></td></tr></tbody></table>

evaluation of blood lactate and plasma insulin during high-intensity exercise by antecubital vein catheterization

Die Messung der endokrinen und metabolischen Marker während der körperlichen Aktivität ist von Bedeutung für das Verständnis der physiologischen Auswirkungen der verschiedenen Übung Modalitäten. Während einige Übung Modalitäten (z. B. hoher Intensität Intervall Übung), Blut Metaboliten und Hormonspiegel ändern in kurzen Zeiträumen. In der vorliegenden Studie beschreiben wir eine Methode, um die Antecubital Vene Katheterisierung ermöglicht die Sammlung von mehreren Blutproben während des Trainings. Insulin und venösen Laktat-Konzentrationen wurden während des intensiven Trainings durch die Anwendung des beschriebenen Verfahrens gemessen. Die Übung bestand aus drei 30 s Anfälle von hoch-intensives Training durch 4 min Erholung getrennt. Nach der letzten Erholungsphase wurde ein Wingate-Test durchgeführt. Blutproben aus der Antecubital Vene wurden vor und nach jedem 30 s-Anfall und vor und nach dem Wingate-Test erhalten. Dadurch war es möglich, die Plasma-Insulin und venöses Blut Laktat Variationen während des Trainings zu bewerten.

Als repräsentative Ergebnisse zu erzielen haben wir dieses Protokoll Blut Laktat und Insulin Variation in sechs Fächern während einer HIIE in einem Fahrradergometer gemessen. Spitzenleistung und Durchschnittsleistung für den Wingate-Test wurden 732 ± 120 und 549 ± 84 Watt, beziehungsweise. Durchschnittsleistung wurde verwendet, um die Ausgangsleistung (494 ± 75 Watt) während die 30 s-Anfälle von den HIIE zu berechnen.
Abbildung 2 zeigt, dass Plasma-Insulin und Blut-Laktat-Variation während der HIIE. Plasma-Insulin (Abbildung 2A) verringerte sich während der ersten und dritten 30 s Kämpfe aber stieg im zweiten Kampf und die Erholungsphasen. Auf der anderen Seite Plasma Laktat (Abb. 2 b) während der zweiten und dritten 30 s Anfälle verringert, aber während der ersten 30 s Kampf und die Erholungsphasen erhöht.
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56890/56890fig1.jpg"> Abbildung 1: Übung Protokoll. Rote Pfeile zeigen die Zeit der Blutentnahme Proben. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56890/56890fig1large.jpg" target="_blank">Bitte klicken Sie hier für eine größere Version dieser Figur.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56890/56890fig2.jpg"> Abbildung 2: experimentelle Ergebnisse. (A) Plasma Insulin (UlU/mL). (B) Blut Laktat (Mmol/L). Jede Farbe steht ein anderes Thema. <a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56890/56890fig2large.jpg" target="_blank">Bitte klicken Sie hier für eine größere Version dieser Figur.</a>

Watch this Scientific Journal Video about Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization at JoVE.com

Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization

Hier präsentieren wir ein Protokoll, um mehrere Blutproben aus der Antecubital Vene während hochintensivem Intervalltraining zu erhalten. Dieses Protokoll kann nützlich sein für die Messung von Blut Metaboliten und endokrine Marker während der Übung.

Auswertung der Blut-Laktat und Plasma-Insulin im hoch-intensives Training von Antecubital Vene Katheterisierung

The measurement of metabolic and endocrinal markers during physical activity is of relevance to understanding the physiological implications of different ...

evaluation-blood-lactate-plasma-insulin-during-high-intensity

Nanyang Technological University

Research

JoVE Journal

Immunology and Infection

6.4K Aufrufe.  Kyoto University. Hier präsentieren wir ein Protokoll, um mehrere Blutproben aus der Antecubital Vene während hochintensivem Intervalltraining zu erhalten. Dieses Protokoll kann nützlich sein für die Messung von Blut Metaboliten und endokrine Marker während der Übung.

Serie: Evaluation of Blood Lactate and Plasma Insulin During High-intensity Exercise by Antecubital Vein Catheterization

Hyperinsulinemic-Euglycemic Clamp in the Conscious Rat

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, T2D results in elevated blood glucose levels. The "gold-standard" in assessing insulin sensitivity is a hyperinsulinemic-euglycemic clamp or insulin clamp. In this procedure, insulin is infused at a constant rate resulting in a drop in blood glucose. To maintain blood glucose at a constant level, exogenous glucose (D50) is infused into the venous circulation. The amount of glucose infused to maintain homeostasis is indicative of insulin sensitivity. Here, we show the basic clamp procedure in the chronically catheterized, unrestrained, conscious rat. This model allows blood to be collected with minimal stress to the animal. Following the induction of anesthesia, a midline incision is made and the left common carotid artery and right jugular vein are catheterized. Inserted catheters are flushed with heparinized saline, then exteriorized and secured. Animals are allowed to recover for 4-5 days prior to experiments, with weight gain monitored daily. Only those animals who regain weight to pre-surgery levels are used for experiments. On the day of the experiment, rats are fasted and connected to pumps containing insulin and D50. Baseline glucose is assessed from the arterial line and used a benchmark throughout the experiment (euglycemia). Following this, insulin is infused at a constant rate into the venous circulation. To match the drop in blood glucose, D50 is infused. If the rate of D50 infusion is greater than the rate of uptake, a rise in glucose will occur. Similarly, if the rate is insufficient to match whole body glucose uptake, a drop will occur. Titration of glucose continues until stable glucose readings are achieved. Glucose levels and glucose infusion rates during this stable period are recorded and reported. Results provide an index of whole body insulin sensitivity. The technique can be refined to meet specific experimental requirements. It is further enhanced by the use of radioactive tracers that can determine tissue specific insulin-stimulated glucose uptake as well as whole body glucose turnover.

The hyperinsulinemic-euglycemic clamp is the "gold standard" for the assessment of insulin action. Insulin is infused at a constant rate stimulating glucose uptake. The amount of exogenous glucose infused to counter this drop is indicative of insulin sensitivity. Here the procedure is performed on a conscious, unrestrained rat.

Hyperinsulinämische-euglykämische Clamp in der Conscious Rat

Type 2 diabetes (T2D) is rapidly rising in prevalence. Characterized by either inadequate insulin production or the inability to utilize insulin produced, ...

Forschung

Medizin

Finger-stick Blood Sampling Methodology for the Determination of Exercise-induced Lymphocyte Apoptosis

Exercise is a physiological stimulus capable of inducing apoptosis in immune cells. To date, various limitations have been identified with the measurement of this phenomenon, particularly relating to the amount of time required to isolate and treat a blood sample prior to the assessment of cell death. Because of this, it is difficult to determine whether reported increases in immune cell apoptosis can be contributed to the actual effect of exercise on the system, or are a reflection of the time and processing necessary to eventually obtain this measurement. In this article we demonstrate a rapid and minimally invasive procedure for the analysis of exercise-induced lymphocyte apoptosis. Unlike other techniques, whole blood is added to an antibody panel immediately upon obtaining a sample. Following the incubation period, red blood cells are lysed and samples are ready to be analyzed. The use of a finger-stick sampling procedure reduces the volume of blood required, and minimizes the discomfort to subjects.

Exercise is capable of inducing apoptosis in immune cells. There are various measurement limitations, particularly relating to the amount of time required to isolate and treat a blood sample prior to the assessment. Demonstrated is a rapid and minimally invasive procedure for the analysis of exercise-induced lymphocyte apoptosis.

Finger-Stick Blutprobenentnahme Methodik zur Bestimmung der Übung-induzierte Lymphozyten Apoptose

Exercise is a physiological stimulus capable of inducing apoptosis in immune cells. To date, various limitations have been identified with the measurement ...

Immunologie und Infektion

Hyperinsulinemic-euglycemic Clamps in Conscious, Unrestrained Mice

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold standard" method for assessing insulin action in vivo. During an insulin clamp, hyperinsulinemia is achieved by a constant insulin infusion. Euglycemia is maintained via a concomitant glucose infusion at a variable rate. This variable glucose infusion rate (GIR) is determined by measuring blood glucose at brief intervals throughout the experiment and adjusting the GIR accordingly. The GIR is indicative of whole-body insulin action, as mice with enhanced insulin action require a greater GIR. The insulin clamp can incorporate administration of isotopic 2[14C]deoxyglucose to assess tissue-specific glucose uptake and [3-3H]glucose to assess the ability of insulin to suppress the rate of endogenous glucose appearance (endoRa), a marker of hepatic glucose production, and to stimulate the rate of whole-body glucose disappearance (Rd).
The miniaturization of the insulin clamp for use in genetic mouse models of metabolic disease has led to significant advances in diabetes research. Methods for performing insulin clamps vary between laboratories. It is important to note that the manner in which an insulin clamp is performed can significantly affect the results obtained. We have published a comprehensive assessment of different approaches to performing insulin clamps in conscious mice1 as well as an evaluation of the metabolic response of four commonly used inbred mouse strains using various clamp techniques2. Here we present a protocol for performing insulin clamps on conscious, unrestrained mice developed by the Vanderbilt Mouse Metabolic Phenotyping Center (MMPC; URL: www.mc.vanderbilt.edu/mmpc). This includes a description of the method for implanting catheters used during the insulin clamp. The protocol employed by the Vanderbilt MMPC utilizes a unique two-catheter system3. One catheter is inserted into the jugular vein for infusions. A second catheter is inserted into the carotid artery, which allows for blood sampling without the need to restrain or handle the mouse. This technique provides a significant advantage to the most common method for obtaining blood samples during insulin clamps which is to sample from the severed tip of the tail. Unlike this latter method, sampling from an arterial catheter is not stressful to the mouse1. We also describe methods for using isotopic tracer infusions to assess tissue-specific insulin action. We also provide guidelines for the appropriate presentation of results obtained from insulin clamps.

The hyperinsulinemic-euglycemic clamp, or insulin clamp, is the gold standard for assessing insulin action in vivo. A method for performing insulin clamps in mice is described. This includes a method for arterial catheterization that permits experiments to be performed in conscious, unrestrained mice with minimal stress.

Hyperinsulinämische-euglykämische Schellen in dem Bewusstsein, Unrestrained Mäuse

Type 2 diabetes is characterized by a defect in insulin action. The hyperinsulinemic-euglycemic clamp, or insulin clamp, is widely considered the "gold ...

Surgical Placement of Catheters for Long-term Cardiovascular Exercise Testing in Swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early cardiopulmonary dysfunction is difficult to diagnose, particularly in animal models, as cardiopulmonary function is often measured invasively, requiring anesthesia. As many anesthetic agents are cardiodepressive, subtle changes in cardiovascular function may be masked. In contrast, chronic instrumentation allows for measurement of cardiopulmonary function in the awake state, so that measurements can be obtained under quiet resting conditions, without the effects of anesthesia and acute surgical trauma. Furthermore, when animals are properly trained, measurements can also be obtained during graded treadmill exercise.
Flow probes are placed around the aorta or pulmonary artery for measurement of cardiac output and around the left anterior descending coronary artery for measurement of coronary blood flow. Fluid-filled catheters are implanted in the aorta, pulmonary artery, left atrium, left ventricle and right ventricle for pressure measurement and blood sampling. In addition, a 20 G&#160;catheter is positioned in the anterior interventricular vein to allow coronary venous blood sampling.
After a week of recovery, swine are placed on a motor-driven treadmill, the catheters are connected to pressure and flow meters, and swine are subjected to a five-stage progressive exercise protocol, with each stage lasting 3 min. Hemodynamic signals are continuously recorded and blood samples are taken during the last 30 sec of each exercise stage.
The major advantage of studying chronically instrumented animals is that it allows serial assessment of cardiopulmonary function, not only at rest but also during physical stress such as exercise. Moreover, cardiopulmonary function can be assessed repeatedly during disease development and during chronic treatment, thereby increasing statistical power and hence limiting the number of animals required for a study.

Here we present a protocol to assess cardiopulmonary function in awake swine, at rest and during graded treadmill exercise. Chronic instrumentation allows for repeated hemodynamic measurements uninfluenced by cardiodepressive anesthetic agents.

Chirurgische Platzierung von Kathetern für die langfristige Herz-Kreislauf und Sportmedizin in Swine

This protocol describes the surgical procedure to chronically instrument swine and the procedure to exercise swine on a motor-driven treadmill. Early ...

Determining the Contribution of the Energy Systems During Exercise

One of the most important aspects of the metabolic demand is the relative contribution of the energy systems to the total energy required for a given physical activity. Although some sports are relatively easy to be reproduced in a laboratory (e.g., running and cycling), a number of sports are much more difficult to be reproduced and studied in controlled situations. This method presents how to assess the differential contribution of the energy systems in sports that are difficult to mimic in controlled laboratory conditions. The concepts shown here can be adapted to virtually any sport.
The following physiologic variables will be needed: rest oxygen consumption, exercise oxygen consumption, post-exercise oxygen consumption, rest plasma lactate concentration and post-exercise plasma peak lactate. To calculate the contribution of the aerobic metabolism, you will need the oxygen consumption at rest and during the exercise. By using the trapezoidal method, calculate the area under the curve of oxygen consumption during exercise, subtracting the area corresponding to the rest oxygen consumption. To calculate the contribution of the alactic anaerobic metabolism, the post-exercise oxygen consumption curve has to be adjusted to a mono or a bi-exponential model (chosen by the one that best fits). Then, use the terms of the fitted equation to calculate anaerobic alactic metabolism, as follows: ATP-CP metabolism = A1 (mL . s-1) x t1 (s). Finally, to calculate the contribution of the lactic anaerobic system, multiply peak plasma lactate by 3 and by the athlete&#x2019;s body mass (the result in mL is then converted to L and into kJ).
The method can be used for both continuous and intermittent exercise. This is a very interesting approach as it can be adapted to exercises and sports that are difficult to be mimicked in controlled environments. Also, this is the only available method capable of distinguishing the contribution of three different energy systems. Thus, the method allows the study of sports with great similarity to real situations, providing desirable ecological validity to the study.

This protocol allows researchers focused on exercise and sports sciences to determine the relative contribution of three different energy systems to the total energy expenditure during a large variety of exercises.

Bestimmung des Beitrages der Energy Systems während des Trainings

One of the most important aspects of the metabolic demand is the relative contribution of the energy systems to the total energy required for a given ...

Biologie

Combined Intravital Microscopy and Contrast-enhanced Ultrasonography of the Mouse Hindlimb to Study Insulin-induced Vasodilation and Muscle Perfusion

It has been demonstrated that insulin's vascular actions contribute to regulation of insulin sensitivity. Insulin's effects on muscle perfusion regulate postprandial delivery of nutrients and hormones to insulin-sensitive tissues. We here describe a technique for combining intravital microscopy (IVM) and contrast-enhanced ultrasonography (CEUS) of the adductor compartment of the mouse hindlimb to simultaneously visualize muscle resistance arteries and perfusion of the microcirculation in vivo. Simultaneously assessing insulin's effect at multiple levels of the vascular tree is important to study relationships between insulin's multiple vasoactive effects and muscle perfusion. Experiments in this study were performed in mice. First, the tail vein cannula is inserted for the infusion of anesthesia, vasoactive compounds and ultrasound contrast agent (lipid-encapsulated microbubbles). Second, a small incision is made in the groin area to expose the arterial tree of the adductor muscle compartment. The ultrasound probe is then positioned at the contralateral upper hindlimb to view the muscles in cross-section. To assess baseline parameters, the arterial diameter is assessed and microbubbles are subsequently infused at a constant rate to estimate muscle blood flow and microvascular blood volume (MBV). When applied before and during a hyperinsulinemic-euglycemic clamp, combined IVM and CEUS allow assessment of insulin-induced changes of arterial diameter, microvascular muscle perfusion and whole-body insulin sensitivity. Moreover, the temporal relationship between responses of the microcirculation and the resistance arteries to insulin can be quantified. It is also possible to follow-up the mice longitudinally in time, making it a valuable tool to study changes in vascular and whole-body insulin sensitivity.

Insulin-induced vasodilation regulates muscle perfusion and increases the microvascular surface area (microvascular recruitment) available for solute exchange between blood and tissue interstitium. Combined intravital microscopy and contrast-enhanced ultrasonography is presented to simultaneously assess insulin&#39;s action at the larger vessels and the microcirculation in vivo.

Kombiniert intravitalen Mikroskopie und Kontrast-verstärkte Sonographie der Maus Megalosauridae Insulin-induzierte Vasodilatation und Muskel-Durchblutung zu untersuchen

It has been demonstrated that insulin's vascular actions contribute to regulation of insulin sensitivity. Insulin's effects on muscle perfusion regulate ...

Experimental Protocol of a Three-minute, All-out Arm Crank Exercise Test in Spinal-cord Injured and Able-bodied Individuals

Reliable exercise protocols are required to test changes in exercise performance in elite athletes. Performance improvements in these athletes may be small; therefore, sensitive tools are fundamental to exercise physiology. There are currently many exercise tests that allow for the examination of exercise capacity in able-bodied athletes, with protocols mainly for lower-body or whole-body exercise. There is a trend to test athletes in a sport-specific setting that closely resembles the actions that the participants are used to performing. Only a few protocols test short-term, high-intensity exercise capacity in participants with an impairment of the lower body. Most of these protocols are very sport-specific and are not applicable to a wide range of athletes. One well-known test protocol is the 30 s Wingate test, which is well-established in cycling and in arm crank exercise testing. This test analyzes high-intensity exercise performance over a 30 s time duration. In order to monitor exercise performance over a longer duration, a different method was modified for application to the upper body. The 3 min, all-out arm crank ergometer test allows athletes to be tested in a manner specific to 1,500 m wheelchair racing (in terms of exercise duration), as well as to upper body exercises such as rowing or hand-cycling. In order to increase the reliability with identical test conditions, it is crucial to precisely replicate settings such as the resistance (i.e., torque factor) and the position of the participants (i.e., the height of the crank, the distance between the crank and the participant, and the fixation of the participant). Another important issue concerns the beginning of the exercise test. Fixed revolutions per minute are required to standardize the test conditions for the start of the exercise test. This exercise protocol shows the importance of accurate operations to reproduce identical test conditions and settings.

We present a protocol to test the aerobic and anaerobic power of the upper body muscles over a duration of 3 min in able-bodied as well as in paraplegic and tetraplegic individuals. The protocol presents specific modifications in its application for upper-body exercise in individuals with or without disability.

Experimentelles Protokoll eines dreiminütigen, all-out Arm-Kurbel-Übungstests in der Wirbelsäule Verletzte und kräftige Einzelpersonen

Reliable exercise protocols are required to test changes in exercise performance in elite athletes. Performance improvements in these athletes may be ...

Postconditioning with Lactate-enriched Blood for Cardioprotection in ST-segment Elevation Myocardial Infarction

The beneficial effects of reperfusion therapy for ST-segment elevation myocardial infarction (STEMI) is attenuated by reperfusion injury. No approach has been proven successful in preventing this injury in the clinical setting to date. Meanwhile, a novel approach for cardioprotection in patients with STEMI, i.e., postconditioning with lactate-enriched blood (PCLeB), has recently been reported. PCLeB is a modification of the original protocol of postconditioning, aimed at increasing the delay in the recovery from tissue acidosis produced during ischemia. This was sought to achieve controlled reperfusion with tissue oxygenation and minimal lactate washout. In this modified postconditioning protocol, the duration of each brief reperfusion is gradually increased in a stepwise manner from 10 to 60 s. Each brief ischemic period lasts for 60 s. At the end of each brief reperfusion, injection of lactated Ringer&#39;s solution (20&#8211;30 mL) is performed directly into the culprit coronary artery immediately before the balloon inflation and the balloon is quickly inflated at the lesion site, so that the lactate is trapped inside the ischemic myocardium during each brief repetitive ischemic period. After seven cycles of balloon inflation and deflation, full reperfusion is performed. Stenting is performed thereafter, and the percutaneous coronary intervention is completed. Excellent in-hospital and 6 month outcomes in a limited number of patients with STEMI treated using PCLeB have already been reported. This method article provides a detailed description of each step of the PCLeB procedures.

Herein, we present a protocol for postconditioning with lactate-enriched blood, which is a novel approach for cardioprotection. This protocol comprises&#160;intermittent reperfusion and timely coronary injections of lactated Ringer&#39;s solution. This can be applied to ST-segment elevation myocardial infarction patients who undergo primary percutaneous coronary intervention.

Nachkonditionierung mit laktatangereichertem Blut für Kardioprotektion im ST-Segment Elevation Myokardinfarkt

The beneficial effects of reperfusion therapy for ST-segment elevation myocardial infarction (STEMI) is attenuated by reperfusion injury. No approach has ...

Measurement of Myocardial Lactate Production for Diagnosis of Coronary Microvascular Spasm

In about a quarter of patients with angina and non-obstructive coronary arteries, no epicardial spasm is noted on coronary arteriography during an angina attack. Since the pressure-rate product is almost identical at rest and the onset of attack in those patients, the decrease in coronary blood flow rather than increased myocardial oxygen consumption is likely to explain myocardial ischemia, indicating a substantial involvement with coronary microvascular spasm (MVS). Myocardial lactate production, which could be defined as a negative myocardial lactate extraction ratio (ratio of the coronary arterial-venous difference in lactate concentration to arterial concentration), is considered indicative of objective evidence to support the emerging myocardial ischemia. Thus, monitoring of the myocardial lactate production and the emergence of chest pain and ischemic electrocardiographic changes during acetylcholine (ACh) provocation testing is of significant value for detecting the entity of MVS. Practically, 1 min after incremental doses of ACh (20, 50, and 100 &#956;g) are administered into the left coronary artery (LCA), paired samples of 1 mL of blood are collected from the LCA ostium and coronary sinus for measurement of lactate concentration by a calibrated automatic lactate analyzer. Then, the development of MVS could be confirmed by negative myocardial lactate extraction ratio despite the absence of angiographically demonstrable epicardial coronary spasm or before its occurrence throughout ACh provocation testing. In conclusion, assessment of myocardial lactate production is essential and valuable for the diagnosis of MVS.

Myocardial lactate production (coronary arterial-venous difference in serum lactate level) during coronary spasm provocation testing is considered as a highly sensitive marker that reflects acetylcholine-induced myocardial ischemia due to microvascular spasm. This article presents the procedures to assess myocardial lactate production for the diagnosis of coronary microvascular spasm.

Messung der myokardialen Laktatproduktion zur Diagnose des koronaren mikrovaskulären Spasmus

In about a quarter of patients with angina and non-obstructive coronary arteries, no epicardial spasm is noted on coronary arteriography during an angina ...

Bewertung des Krafttrainingsvolumens und seiner Auswirkungen auf die Insulinsensitivität bei Adipositas

Randomized Controlled Trial to Study the Acute Effects of Strength Exercise on Insulin Sensitivity in Obese Adults

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of sets) have not been studied thoroughly. Although it is intuitive that some SE is better than none, and more is better than some for the improvement of IS, high-volume sessions might be challenging for diseased populations to complete, especially obese adults, for whom even a brisk walk can be challenging. This protocol details a randomized clinical trial to assess the acute effects of SE on IS in obese adults. The inclusion criteria are body mass index &#62;30 kg/m2, central obesity (waist circumference &#62;88 cm and &#62;102 cm for women and men, respectively), and age &#62;40 years. Participants will be familiarized with the SE (7 exercises targeting major muscle groups) and then will perform three sessions in a randomized order: session 1 - high-volume session (3 sets/exercise); session 2 - low-volume session (1 set/exercise); session 3 - control session (no exercise). Diet will be controlled the day before and on the day of the sessions. Sessions will be completed at night, and an oral glucose tolerance test will be performed the next morning, from which several indexes of IS will be derived, such as the area under the curve (AUC) of glucose and insulin, the Matsuda index, the Cederholm index, the muscle IS index, and the Gutt index. Based on pilot studies, we expect ~15% improvement in IS (insulin AUC, and Matsuda and Cederholm indexes) after the high-volume session, and ~8% improvement after the low-volume session compared to the control session. This study will benefit individuals who find high-volume SE sessions challenging but still aim to improve their IS by investing 1/3 of their time and effort.

Autor im Rampenlicht: Erforschung der Auswirkungen eines reduzierten Widerstandstrainingsvolumens auf die metabolische Gesundheit 

This study describes a randomized controlled trial protocol aiming at assessing the acute effects of strength exercise volume on insulin sensitivity in obese individuals.

Randomisierte kontrollierte Studie zur Untersuchung der akuten Auswirkungen von Krafttraining auf die Insulinsensitivität bei adipösen Erwachsenen

An acute session of strength exercise (SE) ameliorates insulin sensitivity (IS) for several hours; however, the effects of SE volume (i.e., number of ...