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

著者は、博士 p. Karagiannis ペーパーを読むと彼の技術的なサポートのための教授 t. ほようを認識したいと思います。

ここで説明したすべてのメソッドは、ローカル倫理委員会によって承認されている (同志社大学倫理委員会: 15033)、ヘルシンキ宣言によって設定された基準に厳密に従っていた。すべての参加者からインフォームド コンセントを得た。被験者は男性のレクリエーション スポーツ選手 (年齢、20.1 ± 1.2 年; 重量 69.7 ± 6.2 kg; 176.1 ± 高さ 5.8 cm; 体脂肪 13.6 ± 1.5% と VO2、最大 52.6 ± 6.6 mL/分1/kg1)。
1. 運動プロトコルの確立
注: 本研究は HIIE プロトコルの中に肘静脈から採血して血液ホルモンと代謝物の動態評価の成っていた。
<ol><li>低強度での自転車エルゴメータでのサイクリングの 5 分を暖かいを実行する対象を求めます。ウォーム アップの後、残り 5 分の件名をしましょう。</li>
	<li>下肢エルゴメータで次の HIIE を実行する対象を求める: 30 s 高強度運動、区切られた (図 1)、残りの 4 分の 3 の発作。 注: 30 s 発作べき強度と同額に設定 90% 以前に実行されたウィンゲート テストのパワー平均。ウィンゲート テストの完全な説明の8を他の場所で参照してください。</li>
	<li>30 s 発作の負荷を計算するには、次の数式を使用します。 仕事 = 力 x 距離 (関係式 1) 力 = 質量 x 加速度 (式 2) 仕事 = (質量 x 加速度) x 距離 (式 3) 注: このモデルは作業負荷 (kg) 質量と加速度は 9.81 m/s2です。距離は、車輪の直径 (1.622 m) です。3.7 時間/peal 回転車輪の回転するため、ペダル回転あたりの距離は 3.7 x 1.662 です。さらに、ペダル回転数は 90 回転/分190 の 1 つのペダル距離の乗算するあります。 仕事 = 9.81 x 3.7 x 1.622 x 90 (関係式 4) x 作業負荷 作業負荷 = 仕事/(9.81 x 3.7 x 1.622 x 90) (式 5)</li>
</ol>2. カテーテル
注: チューブと vacutainers ラベルを付けるように事前に時間を失うことを避けるために。
<ol><li>カテーテル検査の選択腕に駆血帯をラップします。これを行うには、選択した arm の背面にあるゴムバンドを配置します。伸縮バンド、腕の周りのバンドで輪を作るし、弾性を結ぶ。</li>
	<li>目に見える肘静脈を選択し、エタノールやアルコールで皮膚を消毒します。</li>
	<li>滅菌のカニューレの先端を皮膚に 15 ° のおおよその角度で静脈に挿入します。 注: から近位遠位方向にカニューレを挿入します。穿刺に成功した場合は、血液のバック フラッシュを観察すべき。</li>
	<li>手でカニューレを固定し、そっと針を削除します。 注: 運動中に針の角度を維持するために綿組織は皮膚と針の間に使用することがあります。この構造体をテープで固定します。</li>
	<li>ヘパリンと最初の血液サンプル (約 2.5 mL) の吸引、バキュテナーを接続します。すぐの後で、40 分のアイス ボックスにそれを置くことによって血液サンプル冷蔵庫で冷やします。血液ガス分析しようとしている場合は、アシスタントを助けを求めます。</li>
	<li>カテーテルにチューブを接続し、血液凝固を避けるために管の長さがフルにヘパリンを注入します。蝶を閉じます。</li>
	<li>テープを使用して、カテーテルを安定させます。この時点でことを確認、被写体の腕移動は血のコレクションと干渉しません。この条件を確認するため針の位置は、任意の動きの影響を受けないことを必ずください。</li>
</ol>3. 血液サンプリング
<ol><li>ヘパリンを削除するには、カテーテルに血液だけがあるまで、バキュテナーを使用します。血液のフラッシュ バックを観測すると、2.5 の手順で説明するようにサンプルを収集します。</li>
	<li>サンプル コレクション後血液凝固を避けるためにカテーテル チューブにヘパリンを再び挿入します。この操作の後ステップ 2.5 は、いくつかのサンプル コレクションの繰り返されるかもしれません。</li>
	<li>最後のサンプルが収集された後慎重にカニュレーション チューブを削除し、アルコールで皮膚をきれいします。可能な限り出血を避けるため、カテーテル検査が作られた場所で滅菌綿を保つために主題を求めます。</li>
	<li>バキュテナー チューブで収集された血液は、慎重に血液の約 1.5 mL を 1.5 mL チューブに渡すし、チューブを氷の上。</li>
	<li>測定血中乳酸乳酸検出器を用いた血の残りから。同じ日に、プラズマの採血 (血液の約 1.5 mL) を取得します。</li>
	<li>血漿分離 4 oC で 5000 × g で 5 分間の血液サンプルを遠心し、新しい 1.5 mL チューブに上清を分離します。</li>
	<li>プラズマを浄化するために遠心分離の 1 つのより多くの時間を繰り返します。</li>
	<li>血漿分離後のホルモンの分析まで-80 oC でサンプルを格納します。</li>
	<li>製造元のあたり (材料の表を参照) をプロトコルとして ELISA キットを使用してインスリンを測定します。</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>

著者が明らかに何もありません。

本研究では肘静脈が我々 だったと血液採取した高インターバル運動中に正常に。この手法の利用例として採血を行った (後に血漿分離) と血中乳酸の血漿インスリンのこの方法の主な利点は、血液サンプルのコレクション要するに期間できるということです。同時に、それはいくつかの制限があります。その使用を許可する限られた運動様式などアシスタントの数、必要し、専門訓練を受けた人材が必要です。腕の血圧が低い結果として低いサンプル ボリュームのリスク。
成功した血液サンプル コレクションの被験者の腕は静的な位置でなければなりません。メソッドは、いくつかの演習の様相にのみ適用できる現在の形で、前腕可動性が減少。ここでは、我々 は、エルゴメータでの運動が実行されますが、変更、それはトレッドミル マシンで使用することができるときにメソッドを適用する方法を説明します。どちらの場合も、採血の経験を持つ人材が必要です。
運動プロトコルに関する解決すべきいくつかの考慮事項があります。いくつかの試験評価 (例えば発作の間のアクティブな回復と) する場合は、テストのための決断の瞬間が重要です。ホルモン変動10,11を避けるためには、別の日に同じ時間ですべての試験を実行することをお勧めします。さらに、代謝は時刻および消費の食事によって影響を受けます。したがって、テストの前に、断食がお勧めします。さらに、再現性を高めるためには、テスト前に被験者による運動プロトコルを習熟期間はお勧めします。この期間、実験科目では、メインのテストに (徐々 に) 同様の運動を実行します。
結論として、我々 は運動中にいくつかの静脈血液サンプルを取得する効果的な方法を説明します。このメソッドは、運動 (例えば、代謝物質、ホルモン、蛋白質、RNA) 中にいくつかの分子を研究するために使用することができますのサンプルを収集するため便利です。

身体活動時代謝と内分泌マーカーの測定は、さまざまな運動療法の生理学的意味を理解するための関連性の高いです。いくつかの運動の様相 (例えば、高強度インターバル トレーニング (HIIE)) 中、血液代謝やホルモンのレベルは比較的短時間1,2,3で変動します。
(すなわちサッカー、バスケット ボール、ラグビー) の多くのスポーツは、低の激しい運動やパッシブ期間4,5で区切られた高強度運動の短い期間で構成されます。HIIE は前述のスポーツとの類似点を提示し、選手6トレーニング法として広く利用されています。さらに、スポーツとの類似性のため HIIE として使用されていますモデル7運動生理学を勉強します。研究目的および物理的な条件の評価に使用頻繁および採血生理学的変数を測定することが欠かせません。ただし、HIIE の中に採血はサンプル コレクション間の短い期間のため技術的に困難することができます。ここで、この問題の解決策として間を取得する血液サンプル肘静脈カテーテル法によって短い期間科目を実行 HIIE の方法を示す.

<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

身体活動時代謝と内分泌マーカーの測定は、さまざまな運動療法の生理学的意味を理解するための関連性の高いです。いくつかの中に運動療法 (例えば、高強度インターバル トレーニング)、血液代謝やホルモンのレベルを変更で短い期間。本研究では運動中にいくつかの血液サンプルのコレクションを可能にする肘静脈カテーテルを入れて下さい方法をについて説明します。記述法の適用によって高強度の運動中インスリンおよび静脈の乳酸濃度を測定しました。運動は、高強度運動回復の 4 分で区切られた 3 つの 30 s 発作から成っていた。最後の回復期間後ウィンゲート テストを行った。前に、各 30 秒試合後と前に、とウィンゲート テスト後、肘の静脈から血液サンプルが得られました。その結果、運動中に血漿インスリンと静脈血乳酸のバリエーションを評価することが可能だった。

代表的な結果としてエルゴで HIIE の中に 6 科目で血の乳酸およびインスリン変動を測定するのにこのプロトコルを使用しました。ピーク電力と平均電力ウィンゲート テスト 732 ± 120 と 549 ± 84 ワットであった。平均電力は、HIIE の 30 s 発作時に出力 (494 ± 75 ワット) を計算していました。
図 2は、血漿インスリンと血乳酸、HIIE 間の変化を示しています。血漿インスリン (図 2 a) は、最初と 3 番目の 30 秒発作時に減少したが、第 2 試合と回復期間の間に増加しました。その一方で、血漿乳酸 (図 2 b) 発作時に、2 番目と 3 番目 30 s、減少したが、まず 30 秒続き、回復期間中に増加しました。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56890/56890fig1.jpg"> 図 1: 運動プロトコル。赤い矢印は、血液サンプルのコレクションの時間を示します。<a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56890/56890fig1large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</a>。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56890/56890fig2.jpg"> 図 2: 実験的結果。(A) インスリン (ウル/mL)。(B) 血液は乳酸 (mmol/L) です。それぞれの色は、別の件名を表します。<a href="https://cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56890/56890fig2large.jpg" target="_blank">この図の拡大版を表示するのにはここをクリックしてください</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

肘静脈から高強度のインターバル トレーニング中にいくつかの血液サンプルを取得するためのプロトコルをご紹介します。このプロトコルは、血液代謝物の測定のため役に立つかもしれません、中に内分泌マーカー運動。

肘静脈カテーテル法による高強度運動時の血中乳酸および血漿インスリンの評価

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 ビュー.  Kyoto University. 肘静脈から高強度のインターバル トレーニング中にいくつかの血液サンプルを取得するためのプロトコルをご紹介します。このプロトコルは、血液代謝物の測定のため役に立つかもしれません、中に内分泌マーカー運動。

ビデオ: 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.

覚醒ラットにおける高インスリン血症、正常血糖クランプ

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

医学

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.

運動誘発性リンパ球のアポトーシスの決定のための指先穿刺の採血方法

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

免疫・感染学

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.

意識、無拘束マウスにおける高インスリン血症、正常血糖クランプ

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.

豚の長期心血管運動テストのためのカテーテルの外科的配置

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.

運動時のエネルギーシステムの貢献を決定する

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 ...

生物学

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.

インスリン依存性血管拡張や筋肉の血流を研究する生体顕微鏡観察とマウスの後肢の造影超音波検査

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.

脊髄損傷および肥満者における3分間の全身アームクランク運動試験の実験プロトコル

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.

STセグメント上昇心筋梗塞における心保護のための乳酸濃縮血液とのポストコンディショニング

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.

冠動脈微小血管痙攣診断のための心筋乳酸産生測定

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

筋力運動量と肥満のインスリン感受性への影響の評価

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.

著者スポットライト:運動量の減少が代謝の健康に与える影響の調査 

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

肥満成人のインスリン感受性に対する筋力運動の急性効果を研究するためのランダム化比較試験

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