This work was supported by a National Imaging Facility Subsidised Access Grant to BJC, a National Health and Medical Research Council of Australia program grant (482800) to KAR and PJB. The authors would like to thank Andrew Arthur, Hasar Hazme and Marie-Claude Gregoire for support in developing this method.

すべてこの研究で説明する手順シドニー現地保健地区、シドニーの動物倫理委員会の大学によって承認され、 実験動物の管理と使用のための NIH ガイドに従った、第8版（2011年）。 1.動物の準備注：このプロトコル雄のdb / dbマウス（BKS.Cg- Dock7 M + / + Lepr のdb / J）で6週齢まで固形飼料および水を随意アクセスの群ハウジングに維持しました。撮影時には、マウスは〜30グラムの重量を量りました。このプロトコルで使用されるすべてのマウスは、10及び14ミリモル/ Lの空腹時血糖値ました。 <ol><li>必要な場合は、マウスを高速。本実施例では、実験手順の前に5時間マウスを絶食。 </li><li>撮像の開始前に所望の薬剤（ 例えば 、薬物、タンパク質、ペプチド）を有するマウスを治療します。この例では、インスリンの皮下注射を投与（3U / kgのヒトインスリン）又は等量のPBS 30分前に撮影を開始します。 </li></ol> 2.ワークフローを設定します注：このプロトコルは、PET / CTスキャナーに実装されました。 CTデータの取得に続いて、最初のPETデータを取得します。 <ol><li> PETの設定： <ol><li> 650 keVの（デフォルト）3.432 NS（デフォルト）の一致タイミングウィンドウで- 18 F、3,600秒に設定し、スキャン時間、及び350 keVのに上下のレベルのエネルギー弁別として同位体を選択します。トレーサー注入後60分 - 期間0 16のフレーム（6×10秒、4×60秒、1×300秒、5×600秒）にヒストグラムリストモードデータ。ズーム1.5で2D-FBPを用いてサイノグラム放射を再構成します。 注：再構成画像16の動的フレーム、128×128×159ボクセルと0.52×0.52×0.796ミリメートル3のボクセルサイズを有する各々から成っていました。 </li></ol></li><li> CTの設定： <ol><li>ために全身CTスキャン、500 A、50キロボルトの電圧、露光時間500ミリ秒と360回転にわたって200個の突起で電流を設定します。 （完全なPET視野範囲をカバーする）3にベッド位置の数、30722048に検出器の視野（FOV）を設定し、4にベッド= 30.234713%位置と検出器ビニング間で重複。 注：CT再構成は、HU較正、バイリニア補間とシェップ - ローガンフィルタとコーン・ビーム断層撮影画像再構成ソフトウエアを用いて行きました。 </li></ol></li><li> 18 F-FDG： <ol><li>十分な18 F-FDGを注文（例えば、0.5 mLの450 MBqの）ローカルプロバイダからは、最初の注射の前に約30分に到着します。動物は、0.1 mLの最終体積で18 F-FDGの約10 MBqのを受け取ることができるように、18 F-FDGをアリコートし、希釈します。 </li></ol></li></ol> 3.イメージングプロトコル<ol><li>無菌状態を維持するために、80％（v / v）のエタノールで誘導チャンバーおよびイメージングベッドを拭います。プラ誘導チャンバ内にマウスをCEおよび酸素中の5％イソフルランで麻酔。 </li><li> 1L / minの流量で - （2％メンテナンス、1.5）イソフルランを送達するために、体温を維持するために電気加熱パッド及び精度気化器のノーズコーンが取り付けられた撮影台上にマウスを置きます。麻酔下ながら乾燥を防ぐために、目に眼軟膏を適用します。 </li><li>呼吸を監視し、麻酔の適切な面が維持されることを保証するために、センサパッド上に腹臥位にマウスを置き。 </li><li>外側尾静脈を拡張するために2分 - 1ための熱パックを使用して、尾を温めます。外側尾静脈に、30ゲージの針を挿入することにより、外側尾静脈にカテーテルを挿入。手術用接着剤で所定の位置に針を固定し、カテーテルを固定します。 </li><li>負荷イメージングは​​、スキャナにベッドとカテーテルが機械の後方からアクセスできるように、機械を介してベッドを移動させます。 </li><li> SYRでの18 F-FDGシリンジにカテーテルを取り付けインゲドライバー。注射して投与される体積（&lt;100μL、10秒かけて注入）の前にシリンジ内のアクティビティに基づいて、正確な18 F-FDGの用量（10 MBqの）を計算します。 </li><li>グルコース取り込みの変動に対する麻酔の影響を最小限に抑えるために、麻酔の誘導および（例えば、30分）、18 F-FDGの注射の間に一定の時間を確保します。 </li><li> PETは、18 F-FDGの注射の直前にスキャン開始。 PETスキャン（3600秒）を終えた後、組織と放射性トレーサー取り込みの共同登録を可能にするためにCTスキャン（〜10分）を行います。 </li><li>開始位置に撮影台を移動させる、床から動物を削除します。 </li><li>この時点で、動物を安楽死させるか、それが回復することができます： <ol><li>安楽死のために、麻酔下でまだながら頸椎脱臼を行い、その後の分析のための関心の臓器を収集します。 </li><li>マウスが回復することができた場合は、加熱パッドの上に単一のハウジング内にマウスを置きますか、加熱ランプの正面です。それは胸骨横臥位を維持するのに十分な意識を取り戻したまでマウスを監視します。マウスは、グループハウジングに戻る前に1時間回復させます。 </li></ol></li></ol> 4. PET画像処理注：画像の再構成は、研究職場ソフトウェアv4.2の中で取得職場ソフトウェアv1.5.0.28と分析を用いて行きました。 <ol><li> CTとPET画像を同時登録及び位置合わせが全て3次元で正確であることを確認してください。 <ol><li> 「ファイル」メニューでは、「フォルダの検索/インポート」を選択し、データを格納しているフォルダを選択します。希望PETとCTデータを選択し、「一般分析」タブをクリックします。 </li><li>ソートCRは、「ソースのデータを指定されているようにPETを「ターゲット」を指定されています。 「ワークフロー」メニューでは、「登録」を選択します。画像が正しく共同登録する調整が必要な場合は、番目のツールを使用E「登録」メニュー。 </li></ol></li><li> 「ワークフロー」メニューでは、「ROI定量化」を選択します。 <ol><li>所望の領域の位置を特定するために「パン」と「画像」タブで「ズーム」機能を使用してください。 「ツール」メニューで、「作成」タブを選択し、絵筆のアイコンをクリックしてください。画像上にROIを描画</li></ol></li><li> 「保存」メニューから「保存ROI定量化」を選択することで、時間放射能曲線を抽出します。 CSVファイルとしてデータを保存します。 </li><li>組織の1cm 3当たりベクレルなどの放射能の取り込みを定量化します。 CSVは、スプレッドシートにファイルをロードすることにより、1cm 3当たりの注射用量率（％ID / cm 3）との比率に値を変換します。 </li></ol> 5.入力機能<ol><li>システムの点広がり関数を補正するために、以前に9に記載されているようreblurredヴァンCittertデコンボリューション法を用いて5回の反復の推定システムのPSFをデコンボリューション。 注：これが原因マウスにおけるヴェナ・キャバの小さなサイズに必要とされます。 </li><li>上述したように、血液入力関数の時間 - 活性曲線を生成するために、ポストデコンボリューション画像を使用します。 </li></ol> 6.キネティックモデル注：FDG二組織コンパートメントモデル（ 図1）血漿入力関数を必要とします。 <ol><li> Input_plasma = Input_blood×（ - 0.191トン + 1.165 0.386 E）：以下の式10を用いてプラズマ入力機能にCSVファイル内の血液入力関数を変換します。 </li><li>運動モデリングツールでは「キネティック」ボタンをクリックしてください。組織および血漿総活性は、「メニュー」から「ロード時間の活性曲線」を選択することで、運動モデリングツールにCSVファイルをインポート数えます。 </li><li> 「モデル」メニューでは2つの組織区画を選択します。 K4の隣のボックスが選択されていないことを確認してくださいそして、VB（血液体積分率）のためのチェックボックスをオフにし、2％の値を入力し、初期フィッティングについては0の値を入力します。 </li><li> 「フィット現在の領域]をクリックします。分散11、12の対象の抽出された領域を修正します。異なる分散時間のためのFDGモデルのカイ二乗値を最小化することによって、これを達成。 </li><li>地域の速度定数（k 1 -k 3）を計算するためにフローティングのVBの値（次のVBのためのボックスにチェックボックス）と最適化された分散値を用いて第2の適合を行います。 K iは =（K 1×K 3）/（K 2 + K 3）のような一定の地域の流入を計算します。 </li></ol>

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The authors have nothing to disclose.

ここで説明するプロトコルは、組織及びマウスにおける後続の代謝への血流からのグルコース取り込みの動態を決定するために、堅牢な非侵襲的方法を表します。 DB / dbマウスは、インスリン抵抗性と関連する介入を調べるために広く使用されている2型糖尿病14の十分に確立された動物モデルです。しかし、以前の研究では唯一の心15?...

本研究の目的は、 インビボで 、マウスにおける特定の組織への血流からのグルコースのリアルタイム取り込みを定量化するために陽電子放射断層撮影/コンピュータ断層撮影法（PET / CT）ベースの方法を開発することでした。これは、細胞内の空間にプラズマから18 F-FDGの取り込みの速度を推定するために、細胞内空間からプラズマへの輸送の速度との速度のグルコース取り込みおよび動態モデリングを測定するための18 F標識フルオロデオキシグルコース（FDG）を使用して達成されました18 F-FDGのリン酸化。 げっ歯類では、18 F-FDGは、多くの癌治療1、腫瘍の進行2および腫瘍代謝3の研究だけでなく、褐色脂肪デポ4、神経炎症5と脳代謝6のイメージングの前臨床評価に使用されてきました</SUP&gt;。 マウス（およびラット）におけるグルコースの組織特異的取り込みを調べるために使用される伝統的な方法は、一般に各組織7における放射能の安楽死、組織収集および測定した3 Hまたは14 Cのいずれかと2-デオキシグルコースの放射性標識を用いた治療を含みます。 PET / CTの使用は、生きた動物で、同時に複数の臓器や地域におけるグルコース取り込みと代謝の非侵襲的な決意することができます。安楽死が必要条件ではないように、さらに、この技術は、長手方向の研究で使用するのに適しています。 2型糖尿病（T2DM）は破壊グルコース代謝およびインスリンに対する減少組織応答に二次高血糖症（インスリン抵抗性）およびインスリン8の十分な量を生成するために、膵臓β細胞のできないことを特徴とします。グルコース取り込みと代謝の動態解析は重要な洞察を提供することができます治療的介入の作用及び効果の機構、ならびに、疾患の進行の高度な監視を可能にします。

<table border="0" cellpadding="0" cellspacing="0" width="654">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PET/CT Scanner</td>
			<td style="width:152px;">Siemens</td>
			<td style="width:119px;">Inveon&#160;</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">18F-FDG</td>
			<td style="width:152px;">PETNET Solutions</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Isoflurane</td>
			<td style="width:152px;">Pharmachem</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">30 guage needle</td>
			<td style="width:152px;">BD</td>
			<td align="right" style="width:119px;">305106</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">PMOD modelling software</td>
			<td style="width:152px;">PMOD Technologies</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">BKS.Cg-Dock7m&#160;+/+&#160;Leprdb/J&#160; mice</td>
			<td style="width:152px;">Jackson Laboratory</td>
			<td style="width:119px;">000642</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Human insulin</td>
			<td style="width:152px;">Sigma-Aldrich</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
</table>

determining glucose metabolism kinetics using 18f-fdg micro-pet/ct

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). Impaired uptake and metabolism of glucose in multiple organ systems due to insulin resistance is a hallmark of type 2 diabetes. The ability of this technique to extract an image-derived input function from the vena cava using an iterative deconvolution method eliminates the requirement of the collection of arterial blood samples. Fitting of tissue and vena cava time activity curves to a two-tissue, three compartment model permits the estimation of kinetic micro-parameters related to the 18F-FDG uptake from the plasma to the intracellular space, the rate of transport from intracellular space to plasma and the rate of 18F-FDG phosphorylation. This methodology allows for multiple measures of glucose uptake and metabolism kinetics in the context of longitudinal studies and also provides insights into the efficacy of therapeutic interventions.

我々は以前グルコース取り込みおよび代謝13の動力学にプラズマアポA-Iレベルの増加の影響を調査するために、db / dbマウスモデルを使用していました。本研究では、リアルタイムでの腓腹筋にプラズマからの18 F-FDGの取り込みを監視するためにPET / CTイメージングの有用性を実証するために、インスリンで処置したdb / dbマウスを使用しました...

Watch this Scientific Journal Video about Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT at JoVE.com

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

使用するグルコース代謝動態を決定します<sup&gt; 18</sup&gt; F-FDGマイクロPET / CT

This paper describes the use of 18F-FDG and micro-PET/CT imaging to determine in vivo glucose metabolism kinetics in mice (and is transferable to rats). ...

determining-glucose-metabolism-kinetics-using-18f-fdg-micro-petct

Research

JoVE Journal

Medicine

Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

13.8K ビュー.  UNSW Australia. This study describes a protocol that uses 18F-FDG and positron emission tomography/computed tomography (PET/CT) imaging, together with kinetic modelling, to quantify the in vivo, real-time uptake of 18F-FDG into tissues.

ビデオ: Determining Glucose Metabolism Kinetics Using 18F-FDG Micro-PET/CT

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

使用したラット腎移植後の急性拒絶反応の非侵襲的イメージング<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

医学

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.

乳幼児の脳代謝と血行動態の非侵襲的光計測

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

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. With the increasing availability of knockout and transgenic derivatives, together with a vast amount of genetic information, mice provide one of the best models to study the molecular mechanisms underlying the pathology and physiology of lung diseases. Inhalation, intranasal instillation, intratracheal instillation, and intratracheal intubation are the most widely used techniques by a number of investigators to administer materials of interest to mouse lungs. There are pros and cons for each technique depending on the goals of a study. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented. This technique was applied to administer bleomycin to mouse lungs as a model to study pulmonary fibrosis.

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular disease and for developing therapeutic intervention. Here a noninvasive intratracheal intubation method that can directly deliver exogenous materials to mouse lungs is presented.

マウス肺の病理生理学を研究するための非侵襲的気管内挿管

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to return to normal in a large majority of patients2, which is mainly due to current lack of clinical treatment for acute stroke. This necessitates understanding the physiological effects of cerebral ischemia on brain tissue over time and is a major area of active research. Towards this end, experimental progress has been made using rats as a preclinical model for stroke, particularly, using non-invasive methods such as 18F-fluorodeoxyglucose (FDG) coupled with Positron Emission Tomography (PET) imaging3,10,17. Here we present a strategy for inducing cerebral ischemia in rats by middle cerebral artery occlusion (MCAO) that mimics focal cerebral ischemia in humans, and imaging its effects over 24 hr using FDG-PET coupled with X-ray computed tomography (CT) with an Albira PET-CT instrument. A VOI template atlas was subsequently fused to the cerebral rat data to enable a unbiased analysis of the brain and its sub-regions4. In addition, a method for 3D visualization of the FDG-PET-CT time course is presented. In summary, we present a detailed protocol for initiating, quantifying, and visualizing an induced ischemic stroke event in a living Sprague-Dawley rat in three dimensions using FDG-PET.

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Brain damage resulting from cerebral ischemia may be non-invasively imaged and studied in rats using pre-clinical positron emission tomography coupled with the injectable radioactive probe, 18F-fluorodeoxyglucose. Further, the use of modern software tools that include volume of interest (VOI) brain templates dramatically increase the quantitative information gleaned from these studies.

と陽電子放射断層撮影法を用いた非侵襲的イメージングとリビングラットにおける脳虚血の分析<sup&gt; 18</sup&gt; F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

Characterization of Metabolic Status in Nonhuman Primates with the Intravenous Glucose Tolerance Test

The intravenous glucose tolerance test (IVGTT) plays a key role in the characterization of glucose homeostasis. When taken together with serum biochemical profiles, inclusive of blood glucose levels in both the fed and fasted state, HbA1c, insulin levels, clinical history of diet, body composition, and body weight status, an assessment of normal and abnormal glycemic control can be made. Interpretation of an IVGTT is done through measurement of changes in glucose and insulin levels over time in relation to the dextrose challenge. Critical components to be considered are: peak glucose and insulin levels reached in relation to T0 (end of glucose infusion), the glucose clearance rate K derived from the slope of rapid glucose clearance in the first 20 min (T1 to T20), the time to return to glucose baseline, and the area under the curve (AUC). These IVGTT measures will show characteristic changes as glucose homeostasis moves from a healthy to a diseased metabolic state5. Herein we will describe the characterization of nonhuman primates (Rhesus and Cynomolgus macaques), which are the most relevant animal model of Type II diabetes (T2D) in humans and the IVGTT and clinical profiles of these animals from a lean healthy, to obese dysmetabolic, and T2D state 8, 10, 11.

The goal of this protocol is to present a standard method to perform intravenous glucose tolerance tests (IVGTTs) to assess glycemic control in nonhuman primates and assess their metabolic status from healthy to dysmetabolic.

静脈内グルコース負荷試験とヒト以外の霊長類における代謝状態のキャラクタリゼーション

The intravenous glucose tolerance test (IVGTT) plays a key role in the characterization of glucose homeostasis. When taken together with serum biochemical ...

An Organotypic High Throughput System for Characterization of Drug Sensitivity of Primary Multiple Myeloma Cells

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and heterogeneity of patient-derived multiple myeloma (MM) cells. This approach consists in seeding primary MM cells freshly extracted from bone marrow aspirates into microfluidic chambers implemented in multi-well plates, each consisting of a reconstruction of the bone marrow microenvironment, including extracellular matrix (collagen or basement membrane matrix) and stroma (patient-derived mesenchymal stem cells) or human-derived endothelial cells (HUVECs). The chambers are drugged with different agents and concentrations, and are imaged sequentially for 96 hr through bright field microscopy, in a motorized microscope equipped with a digital camera. Digital image analysis software detects live and dead cells from presence or absence of membrane motion, and generates curves of change in viability as a function of drug concentration and exposure time. We use a computational model to determine the parameters of chemosensitivity of the tumor population to each drug, as well as the number of sub-populations present as a measure of tumor heterogeneity. These patient-tailored models can then be used to simulate therapeutic regimens and estimate clinical response.

We describe a high-throughput drug sensitivity assay for primary multiple myeloma cells. It consists of a reconstruction of the bone marrow microenvironment (including extracellular matrix and stroma) in multi-well plates, and a non-invasive method for longitudinal quantification of cell viability.

一次多発性骨髄腫細胞の薬剤感受性の特徴付けのための器官のハイスループットシステム

In this work we describe a novel approach that combines ex vivo drug sensitivity assays and digital image analysis to estimate chemosensitivity and ...

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test (OGTT) and Insulin Tolerance Test (ITT)

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to insulin-stimulated glucose uptake and a gross decompensation of systemic glucose metabolism. Despite considerable progress in the understanding of glucose metabolism, the molecular mechanisms of its regulation in health and disease remain under-investigated, while novel approaches to prevent and treat diabetes are urgently needed. Diet derived glucose stimulates the pancreatic secretion of insulin, which serves as the principal regulator of cellular anabolic processes during the fed-state and thus balances blood glucose levels to maintain systemic energy status. Chronic overfeeding triggers meta-inflammation, which leads to alterations in peripheral insulin receptor-associated signaling and thus reduces the sensitivity to insulin-mediated glucose disposal. These events ultimately result in elevated fasting glucose and insulin levels as well as a reduction in glucose tolerance, which in turn serve as important indicators of insulin resistance. Here, we present a protocol for the generation and metabolic characterization of high-fat diet (HFD)-fed mice as a frequently used model of diet-induced insulin resistance. We illustrate in detail the oral glucose tolerance test (OGTT), which monitors the peripheral disposal of an orally administered glucose load and insulin secretion over time. Additionally, we present a protocol for the insulin tolerance test (ITT) to monitor whole-body insulin action. Together, these methods and their downstream applications represent powerful tools to characterize the general metabolic phenotype of mice as well as to specifically assess alterations in glucose metabolism. They may be especially useful in the broad research field of insulin resistance, diabetes and obesity to provide a better understanding of pathogenesis as well as to test the effects of therapeutic interventions.

Study of In Vivo Glucose Metabolism in High-fat Diet-fed Mice Using Oral Glucose Tolerance Test OGTT and Insulin Tolerance Test ITT

The current article describes the generation and metabolic characterization of high-fat diet-fed mice as a model of diet-induced insulin resistance and obesity. It further features detailed protocols to perform the oral glucose tolerance test and the insulin tolerance test, monitoring whole-body alterations of glucose metabolism in vivo.

経口ブドウ糖負荷試験 (OGTT) とインスリン負荷試験 (ITT) を使用して高脂肪食飼育マウスの生体内糖代謝の研究

Obesity represents the most important single risk factor in the pathogenesis of type 2 diabetes, a disease which is characterized by a resistance to ...

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to expend energy has made it a potential target for novel therapies to ameliorate obesity and associated metabolic disorders in humans. Though this tissue has been well studied in small animals, BAT&#39;s thermogenic capacity in humans remains largely unknown due to the difficulties of measuring its volume, activity, and distribution. Identifying and quantifying active human BAT is commonly performed using 18F-Fluorodeoxyglucose (18F-FDG) positron emission tomography and computed tomography (PET/CT) scans following cold-exposure or pharmacological activation. Here we describe a detailed image-analysis approach to quantify total-body human BAT from 18F-FDG PET/CT scans using an open-source software. We demonstrate the drawing of user-specified regions of interest to identify metabolically active adipose tissue while avoiding common non-BAT tissues, to measure BAT volume and activity, and to further characterize its anatomical distribution. Although this rigorous approach is time-consuming, we believe it will ultimately provide a foundation to develop future automated BAT quantification algorithms.

Whole Body and Regional Quantification of Active Human Brown Adipose Tissue Using 18F-FDG PET/CT

Using free, open-source software, we have developed an analytical approach to quantify total and regional brown adipose tissue (BAT) volume and metabolic activity of BAT using 18F-FDG PET/CT.

人間の活動の地域の定量化と全身褐色脂肪組織18F-FDG PET/CT を使用

In endothermic animals, brown adipose tissue (BAT) is activated to produce heat for defending body temperature in response to cold. BAT&#39;s ability to ...

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of disorders of consciousness, disrupted higher brain function, and verbal disturbances. As a consequence of an insufficient ability to communicate, objective evaluations are needed from family members, medical staff, and caregivers. One such evaluation is the assessment of functioning brain areas. Recently, multimodal brain imaging has been used to explore the function of damaged brain areas. [18F]-fluorodeoxyglucose positron emission tomography-computed tomography ([18F]FDG-PET/CT) is a successful tool for examining brain function. However, the assessment of brain glucose metabolism based on [18F]FDG-PET/CT is not standardized and depends on several varying parameters, as well as the patient&#39;s condition. Here, we describe a series of semiquantitative assessment protocols for a region-of-interest (ROI) image analysis using self-produced [18F]FDG tracers in patients with sTBI. The protocol focuses on screening the participants, preparing the [18F]FDG tracer in the hot lab, scheduling the acquisition of [18F]FDG-PET/CT brain images, and measuring glucose metabolism using the ROI analysis from a targeted brain area.

Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury

[18F]-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography is useful for studying glucose metabolism related to brain function. Here, we present a protocol for an [18F]FDG tracer set-up and semiquantitative assessment of the region-of-interest analysis for targeted brain areas associated with clinical manifestations in patients with severe traumatic brain injury.

重症脳損傷患者 [18F] FDG トレーサーを用いた定量的評価

Patients with severe traumatic brain injury (sTBI) have difficulty knowing whether they are accurately expressing their thoughts and emotions because of ...

Visualization of Endogenous Mitophagy Complexes In Situ in Human Pancreatic Beta Cells Utilizing Proximity Ligation Assay

Mitophagy is an essential mitochondrial quality control pathway, which is crucial for pancreatic islet beta cell bioenergetics to fuel glucose-stimulated insulin release. Assessment of mitophagy is challenging and often requires genetic reporters or multiple complementary techniques not easily utilized in tissue samples, such as primary human pancreatic islets. Here we demonstrate a robust approach to visualize and quantify formation of key endogenous mitophagy complexes in primary human pancreatic islets. Utilizing the sensitive proximity ligation assay technique to detect interaction of the mitophagy regulators NRDP1 and USP8, we are able to specifically quantify formation of essential mitophagy complexes in situ. By coupling this approach to counterstaining for the transcription factor PDX1, we can quantify mitophagy complexes, and the factors that can impair mitophagy, specifically within beta cells. The methodology we describe overcomes the need for large quantities of cellular extracts required for other protein-protein interaction studies, such as immunoprecipitation (IP) or mass spectrometry, and is ideal for precious human islet&#160;samples generally not available in sufficient quantities for these approaches. Further, this methodology obviates the need for flow sorting techniques to purify beta cells from a heterogeneous islet population for downstream protein applications. Thus, we describe a valuable protocol for visualization of mitophagy highly compatible for use in heterogeneous and limited cell populations.

This protocol outlines a method for quantitative analysis of mitophagy protein complex formation specifically in beta cells from primary human islet samples. This technique thus allows analysis of mitophagy from limited biological material, which are crucial in precious human pancreatic beta cell samples.

近接リゲーションアッセイを利用したヒト膵臓ベータ細胞における内因性ミトファジー複合体の可視化

Mitophagy is an essential mitochondrial quality control pathway, which is crucial for pancreatic islet beta cell bioenergetics to fuel glucose-stimulated ...

使用するグルコース代謝動態を決定します

要約

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この動画の章

使用したラット腎移植後の急性拒絶反応の非侵襲的イメージング

乳幼児の脳代謝と血行動態の非侵襲的光計測

マウス肺の病理生理学を研究するための非侵襲的気管内挿管

と陽電子放射断層撮影法を用いた非侵襲的イメージングとリビングラットにおける脳虚血の分析

静脈内グルコース負荷試験とヒト以外の霊長類における代謝状態のキャラクタリゼーション

一次多発性骨髄腫細胞の薬剤感受性の特徴付けのための器官のハイスループットシステム

経口ブドウ糖負荷試験 (OGTT) とインスリン負荷試験 (ITT) を使用して高脂肪食飼育マウスの生体内糖代謝の研究

人間の活動の地域の定量化と全身褐色脂肪組織¹⁸F-FDG PET/CT を使用

重症脳損傷患者 [¹⁸F] FDG トレーサーを用いた定量的評価

近接リゲーションアッセイを利用したヒト膵臓ベータ細胞における内因性ミトファジー複合体の可視化