We would like to thank the Vanderbilt University Institute of Imaging Science MRI technologists David Pennell, Leslie McIntosh, and Kristen George-Durrett, and the team of Vanderbilt University Medical Center PET/CT technologists led by Martha D. Shone. This work was supported by the following grants from the NIH: NCATS/NIH UL1 RR024975, NIDDK/NIH R21DK096282, NCI/NIH R25CA136440, and NIBIB/NIH T32EB014841.

The authors declare that they have no competing financial interests or other conflicts of interests.

記載した試験プロトコルは、特定の対象に基づいて自動的にセグメントBATデポ温熱および寒冷活性化の両方のPET / CTを使用するように設計されている。関心のあるこれらの自動的に生成された領域は、その後、同じ被験者のPET / CTスキャンに同時登録されている両方の温熱および寒冷活性化MRIスキャンに適用することができる。我々の知る限り、これは同じ健康成人のボランティアでの温熱と...

による世界的な肥満の著しい上昇、エネルギーバランスを理解することを目的とした研究分野の関心の高まりがある。肥満はそれ公衆衛生1の関心の重要な地域作り、糖尿病、肝臓病、心血管疾患や癌などのコストがかかり、壊滅的な医学的状態を招くおそれがあります。エネルギー消費に対するエネルギー摂取量のバランスを理解するための研究の一領域は、褐色脂肪組織またはBATの研究である。脂肪組織と呼 ​​ばれるが、BATは多くの点2で、より一般的な白色脂肪組織（WAT）とは異なる。白色脂肪細胞の機能は、細胞当たり単一の大きな脂質液胞にトリグリセリドを保存するために、必要なとき血流へのエネルギー源として、これらのトリグリセリドを解放することである。非常に異なるようにして、褐色脂肪細胞の機能は、熱を生成することである。これが発生することで1つのメカニズムは、寒さへの曝露を介して行われます。これはsympathetiの増大を引き起こす今度はBATを活性化し、C神経系の活動、。活性化されると、褐色脂肪細胞は、熱を発生させる。そうするために、それらは、細胞当たり多数の小さな脂質小胞に含まれるトリグリセリドを使用して、脱共役タンパク質1（UCP1）の存在を通じて豊富なミトコンドリアにおいて、エントロピー損失をもたらす、ATPの生産なしに代謝基質にトリグリセリドを変換する発熱など。小さ ​​な脂質液胞に格納トリグリセリドが枯渇しているように、脂肪細胞は、血流3中に存在するグルコースおよびトリグリセリドの両方を占める。 9 - BATを研究への関心が劇的に起因する非震え熱発生、身体のエネルギー消費の調節におけるその役割、およびBATと肥満3との間の潜在的な逆の関係への貢献に、近年増加している。さらに、最近の動物研究は、BATがクリアトリグリセリドおよびグルコースfのに重要な役割を果たして示す特に高脂肪食10,11の摂取後、血流中にデキュー。 15 -しかし、我々はBATについて知っていることのほとんどはBAT 4,9,12の多くのデポが含まれている小型哺乳類、研究の結果である。いくつかの初期の研究にもかかわらず、16 - 18、ヒトでのBATの存在は広く人間のBATの研究に興味が更新された際に、最近まで、年齢とともに減少すると考えられていた。 24 -最近の研究では、BATの比較的少量成人19に続くことを示唆している。 BATを研究するための付加的な制限因子は別として、生検及び組織学的染色から、BATを検出するための現在受け入れられている明確な方法は、18 F-フルオロ（18 F-FDG）ポジトロン放出断層撮影法（PET）であることである。現代のPETスキャナは、典型的には、コンピュータ断層撮影（CT）スキャナと組み合わされる。寒冷曝露によって活性化されると、BATは18を占める</s&gt;アップBATが非アクティブ20,21,23,25である場合に18 F-FDG取り込みの非常に低いレベルと比較して、グルコースの代謝アナログであり、PET画像に見えるようになるF-FDG放射性トレーサー、。 PET-CTスキャナの助けにPET検査の間に取得されたCT画像は、解剖学的情報を提供することにより、高18 F-FDGの取り込みと組織とを区別する。 PET-CTイメージングの使用は、（CTスキャンからの線量は無視できないが、PETから主に）電離放射線への被験体を露出させるため、BAT検出のために望ましくない方法である。 健康な成人ヒトでのBATに関する研究の数が増加しているが、人間のBATの最近の研究は、主に回顧PET-CTに限られていた人間の乳児の死体26,27、すでにのために病院に入院してきた人間の青年、19,25研究その他の理由で27から30、および健康な成人の少数のヒトの研究31から35。 BATに影響を与える可能性がある、病気のある患者集団を研究するとき、子供と後ろ向き研究の両方の研究と課題の一つは、変更された結果の可能性である。グルコースは、BAT 36の好ましい燃料源ではないのでさらに、PET研究は常にアクティブにBATが検出されないことがあり、したがって、BATの存在をunderrepresentことがあります。生物医学イメージングとBATの研究に別の困難は、組織デポの境界を定義するために画像分割を実行することに関連している。現在、ヒトでの研究でBATのセグメンテーションは、多くの場合、手動の画像分割のある程度に依存しているため、BATデポの誤認と同様に、評価者間の変動に対して脆弱である。 そのため、自動化された分割方法と一緒に、WAT分布からBATを区別することができる強力な新しいと研究者を提供するこれらの課題に、信頼性の高い空間マッピング技術のBATを勉強するとOL。磁気共鳴イメージング（MRI）は、識別、空間マッピング、およびBATの体積定量化のための機能を有しており、撮像対象のための放射性用量を含む既存のハイブリッドPET-CTイメージング手法とは異なり、MRIは電離放射線を伴わない、安全に使用することができ繰り返し。 MRIを用いて、BATを特定し、定量する能力は、臨床内分泌学と肥満研究の新しい道の追求に劇的なプラスの影響を与える可能性があります。 WAT 90％の脂肪15,26の上にあるのに対して、マウスおよびヒトの両方におけるBATの前脂肪水MRI（FWMRI）の研究では、BATの脂肪信号フラクション（FSF）が40〜80％の脂肪分の範囲であることを示している、27。したがって、我々は、この定量FWMRIメトリックは、他の定量的なMRI測定基準と組み合わせて、ヒトでのBATデポを視覚化し、定量化するために、将来の研究に使用することができると仮定した。会った上でこれはBATの影響を研究するための強力なツールを使って研究コミュニティを提供する電離放射線を使用することなくabolism及びエネルギー消費。 我々の研究グループは、過去3年間、成人のヒトでのBATを研究してきた。 1成人のヒト対象における疑わBATを調査するMRIの使用についての私たちの最初の公開プレゼンテーションでは、ロングビーチ、カリフォルニア州37における磁気共鳴医学のための国際協会（ISMRM）脂肪水分離ワークショップ2012年2月に発生した。 2ヵ月後、私たちのグループは、オーストラリアのメルボルン38で2012年4月におけるISMRMの第 20 回年次総会で、大人2名で疑われるBATでFSFの値を提示した。ソルトレイクシティ、ユタ州で2013年4月におけるISMRMの第 21回年次総会で一年後、この原稿に記載されているプロトコルは、PET-確認されたのMRI定量化の最初の（私たちの知る限り）公共のプレゼンテーションのために使用された成人ヒト被験者39でBAT。具体的には、previouslことを示す証拠を提示しyの疑いのBATが活性冷及び温熱18 F-FDG PET-CTイメージングの両方を使用して活性化可能BATであることを確認した。 2013年以来、健康な成人被験者の私たちのコホートは、温熱と寒冷活性化条件の下で、MRIとPET / CTの両方で画像化は、褐色脂肪の役割を探る」、最近のワークショップで、2014年2月に示された結果との20以上の被験者に拡大している人間の「NIH NIDDK 40が主催。 BATのROIがアクティブに冷たいと温熱PET-CTに基づく自動セグメント化アルゴリズムを使用して線引きして具体的には、成人で18 F-FDG PET-CTで確認された鎖骨上BATの地域でのFWMRI FSFおよびR 2 *緩和特性を報告したスキャン。最近では、我々は18 F-FDG PET-CTで温度マッピングの結果を発表し、高度なFWMRI温度測定41,42を使用して、成人で、BATを確 ​​認した。 手順は、ここで提示獲得同じテーマのS MRIと18 F-FDG PET-CTスキャンの両方、活性化した冷や温熱条件の両方に暴露した後、それぞれ。活性化した冷や温熱18 F-FDG PET-CTスキャンは、被験者の特定に基づき、関心（ROIを）の自動的にセグメント化されたBAT領域を作成するために使用されている。これらのBATのROIがその後PET-CTにおけるMRIの特性を測定するために共同登録MRIスキャンに適用されるBATを確認した。 このプロトコルの制限は、温かいまたは冷たい刺激のいずれかに被写体を露光する際に使用される空気の温度がすべての教科のために一貫しているということです。各被験者の経験が温かく感じたり冷やしれる温度が異なる可能性があるためこれが制限です。従って、空気の温度は、個々の応答に合わせて調整されている間のトライアルセッションを実行し、次に温熱および寒冷活性化プロトコルの間に、これらの温度を使用することによって、より良好な応答を得ることができるかもしれない褐色脂肪組織から。

<table border="0" cellpadding="0" cellspacing="0" width="1130">
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		<tr height="100">
			<td height="100" style="height:100px;width:363px;">Name of Material/ Equipment</td>
			<td style="width:392px;">Company</td>
			<td style="width:375px;">Catalog Number</td>
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		<tr height="38">
			<td height="38" style="height:39px;width:363px;">MRI</td>
			<td style="width:392px;">Philips</td>
			<td style="width:375px;">Achieva 3T</td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:363px;">MRI Torso-XL coil</td>
			<td style="width:392px;">Philips</td>
			<td style="width:375px;">Philips SENSE XL Torso coil 16-elements</td>
		</tr>
		<tr height="68">
			<td height="68" style="height:68px;width:363px;">MRI X-tend Table</td>
			<td style="width:392px;">X-Tend</td>
			<td style="width:375px;">X-tend table, Acieva 3T compatible</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:363px;">X-tend armsupport</td>
			<td style="width:392px;">X-Tend</td>
			<td style="width:375px;">X-tend, accessories</td>
		</tr>
		<tr height="34">
			<td height="34" style="height:34px;width:363px;">X-tend fabricsling</td>
			<td style="width:392px;">X-Tend</td>
			<td style="width:375px;">X-tend, accessories</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">PET/CT</td>
			<td style="width:392px;">GE</td>
			<td style="width:375px;">Discovery STE</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Portable A/C Unit</td>
			<td style="width:392px;">Soleus Air</td>
			<td style="width:375px;">XL-140, 14000 BTU</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Floor fan</td>
			<td style="width:392px;">Lasko Pedestal Fan</td>
			<td style="width:375px;">2527</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Portable Heater</td>
			<td style="width:392px;">Lasko Ceramic Air</td>
			<td style="width:375px;">5536</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Chair</td>
			<td style="width:392px;">Winco Lifecare Recliner</td>
			<td style="width:375px;">585</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Sublingual Thermometer</td>
			<td style="width:392px;">WelchAllyn</td>
			<td style="width:375px;">SureTemp Plus 690</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Cold vest</td>
			<td style="width:392px;">Polar Products</td>
			<td style="width:375px;">Cool58 #PCVZ</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:39px;width:363px;">Thermal IR Camera</td>
			<td style="width:392px;">FLUKE</td>
			<td style="width:375px;">TIR-125</td>
		</tr>
	</tbody>
</table>

human brown adipose tissue depots automatically segmented by positron emission tomography/computed tomography and registered magnetic resonance images

確実に非侵襲的イメージング法を用いて、他の組織から褐色脂肪組織（BAT）を微分すると、ヒトにおけるBATを研究するための重要なステップである。検出陽電子放出断層撮影/コンピュータ断層撮影（PET-CT）により測定されるBATは、典型的には、冷刺激に対する対象の曝露後の走査は、脂肪組織貯蔵所に注射し、放射性トレーサー18 F-フルオロデオキシ（18 F-FDG）の取り込みによって確認される。脂肪 - 水分離、磁気共鳴イメージング（MRI）は、放射性トレーサーを使用することなく、BATを区別する能力を有する。今日までに、ヒト成人におけるBATのMRIは、コールド活性化PET-CTと同時登録されていない。したがって、このプロトコルは、自動的にその後の共同登録されたために、同じ被験者のMRIスキャンを適用するBATマスクを生成するために、18 F-FDG PET-CTスキャンを使用しています。このアプローチは、手動セグメンテーションすることなく、BATの定量的MRI特性の測定を可能にします。 BATマスクは2のPEから作成されるT-CTスキャン：2時間のいずれかに温熱（TN）（24℃）または低温の活性化（CA）（17℃）の条件に暴露した後。 TNおよびCA PET-CTスキャンは、登録されており、標準化されたPETの取り込みおよびCTハウンズフィールド値のみBATを含むマスクを作成するために使用される。 CAおよびTN MRIスキャンは、同じ被写体に取得し、自動的に定義BATマスク内の定量的なMRIのプロパティを確立するために、PET-CTスキャンに登録されている。このアプローチの利点は、セグメント化が完全に自動化され、活性化されたBAT（PET-CT）を同定するための広く受け入れられている方法に基づいていることである。 BATの定量的MRI特性は、PET-CTに関連した放射線を回避MRI専用のBAT審査の基礎として役立つことができるこのプロトコルを使用して確立。

（CA）は、PET-CTやMRI、同じ被写体にMRIやPET-CTスキャンの両方を取得し、すべてのスキャンに共同登録を実行すると、BATの定量的MRI測定基準の信頼性の高い測定を可能にします。 図1は未処理（TN）暖かく寒さを示しています1被験者からスキャンします。 TNおよびCA PET-CTデータの両方を取得することによって、それは明らかに増加した18 F-FDG取り込みによってコールド活性?...

Watch this Scientific Journal Video about Human Brown Adipose Tissue Depots Automatically Segmented by Positron Emission Tomography/Computed Tomography and Registered Magnetic Resonance Images at JoVE

Human Brown Adipose Tissue Depots Automatically Segmented by Positron Emission Tomography/Computed Tomography and Registered Magnetic Resonance Images

ここに提示された方法は、各々が温熱（24°Cに2時間暴露後にスキャンし、18 F-フルオロデオキシ（18 F-FDG）陽電子放出断層撮影/コンピュータ断層撮影法（PET-CT）および脂肪-水分離、磁気共鳴イメージング（MRI）を使用し）と成人のヒト対象における褐色脂肪組織（BAT）をマッピングするために冷たい条件（17℃）。

人間の褐色脂肪組織デポは自動的に陽電子放射断層撮影/コンピュータ断層撮影法で区分されると磁気共鳴画像登録

Reliably differentiating brown adipose tissue (BAT) from other tissues using a non-invasive imaging method is an important step toward studying BAT in ...

human-brown-adipose-tissue-depots-automatically-segmented-positron

Research

JoVE Journal

Medicine

12.1K ビュー.  Vanderbilt University. ここに提示された方法は、各々が温熱（24°Cに2時間暴露後にスキャンし、18 F-フルオロデオキシ（18 F-FDG）陽電子放出断層撮影/コンピュータ断層撮影法（PET-CT）および脂肪-水分離、磁気共鳴イメージング（MRI）を使用し）と成人のヒト対象における褐色脂肪組織（BAT）をマッピングするために冷たい条件（17℃）。 

ビデオ: Human Brown Adipose Tissue Depots Automatically Segmented by Positron Emission Tomography/Computed Tomography and Registered Magnetic Resonance Images

Basic Surgical Techniques in the G&ouml;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

The emergence of the G&ouml;ttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects the close resemblance of these animals to human anatomy and physiology 1-6.The size of the G&ouml;ttingen minipig permits the use of surgical equipment and advanced imaging modalities similar to those used in humans 6-8. The aim of this instructional video is to increase the awareness on the value of minipigs in biomedical research, by demonstrating how to perform tracheal intubation, transurethral bladder catheterization, femoral artery and vein catheterization, as well as transcardial perfusion. 
Endotracheal Intubation should be performed whenever a minipig undergoes general anesthesia, because it maintains a patent airway, permits assisted ventilation and protects the airways from aspirates. Transurethral bladder catheterization can provide useful information about about hydration state as well as renal and cardiovascular function during long surgical procedures. Furthermore, urinary catheterization can prevent contamination of delicate medico-technical equipment and painful bladder extension which may harm the animal and unnecessarily influence the experiment due to increased vagal tone and altered physiological parameters. Arterial and venous catheterization is useful for obtaining repeated blood samples and monitoring various physiological parameters. Catheterization of femoral vessels is preferable to catheterization of the neck vessels for ease of access, when performing experiments involving frame-based stereotaxic neurosurgery and brain imaging. When performing vessel catheterization in survival studies, strict aseptic technique must be employed to avoid infections6. Transcardial perfusion is the most effective fixation method, and yields preeminent results when preparing minipig organs for histology and histochemistry2,9. For more information about anesthesia, surgery and experimental techniques in swine in general we refer to Swindle 2007. Supplementary information about premedication and induction of anesthesia, assisted ventilation, analgesia, pre- and postoperative care of G&ouml;ttingen minipigs are available via the internet at <a href="http://www.minipigs.com">http://www.minipigs.com</a>10. For extensive information about porcine anatomy we refer to Nickel et al. Vol. 1-511.

Basic Surgical Techniques in the G&amp;#246;ttingen Minipig: Intubation, Bladder Catheterization, Femoral Vessel Catheterization, and Transcardial Perfusion

The use of domestic and miniature pigs in science has increased significantly in recent years. By demonstrating how to perform intubation, transurethral bladder catheterization, femoral artery and vein catheterization, as well as transcardial perfusion, we aim to further increase the value of G&ouml;ttingen minipigs in biomedical research.

ゲッティンゲンミニ豚の基本的な手術テクニック：挿管、膀胱へのカテーテル挿入、大腿血管カテーテル法、およびTranscardial灌流

The emergence of the G&ouml;ttingen minipig in research of topics such as neuroscience, toxicology, diabetes, obesity, and experimental surgery reflects ...

医学

Quantification of Atherosclerotic Plaque Activity and Vascular Inflammation using [18-F] Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography (FDG-PET/CT)

Conventional non-invasive imaging modalities of atherosclerosis such as coronary artery calcium (CAC)1 and carotid intimal medial thickness (C-IMT)2 provide information about the burden of disease. However, despite multiple validation studies of CAC3-5, and C-IMT2,6, these modalities do not accurately assess plaque characteristics7,8, and the composition and inflammatory state of the plaque determine its stability and, therefore, the risk of clinical events9-13.
[18F]-2-fluoro-2-deoxy-D-glucose (FDG) imaging using positron-emission tomography (PET)/computed tomography (CT) has been extensively studied in oncologic metabolism14,15. Studies using animal models and immunohistochemistry in humans show that FDG-PET/CT is exquisitely sensitive for detecting macrophage activity16, an important source of cellular inflammation in vessel walls. More recently, we17,18 and others have shown that FDG-PET/CT enables highly precise, novel measurements of inflammatory activity of activity of atherosclerotic plaques in large and medium-sized arteries9,16,19,20. FDG-PET/CT studies have many advantages over other imaging modalities: 1) high contrast resolution; 2) quantification of plaque volume and metabolic activity allowing for multi-modal atherosclerotic plaque quantification; 3) dynamic, real-time, in vivo imaging; 4) minimal operator dependence. Finally, vascular inflammation detected by FDG-PET/CT has been shown to predict cardiovascular (CV) events independent of traditional risk factors21,22 and is also highly associated with overall burden of atherosclerosis23. Plaque activity by FDG-PET/CT is modulated by known beneficial CV interventions such as short term (12 week) statin therapy24 as well as longer term therapeutic lifestyle changes (16 months)25.
The current methodology for quantification of FDG uptake in atherosclerotic plaque involves measurement of the standardized uptake value (SUV) of an artery of interest and of the venous blood pool in order to calculate a target to background ratio (TBR), which is calculated by dividing the arterial SUV by the venous blood pool SUV. This method has shown to represent a stable, reproducible phenotype over time, has a high sensitivity for detection of vascular inflammation, and also has high inter-and intra-reader reliability26. Here we present our methodology for patient preparation, image acquisition, and quantification of atherosclerotic plaque activity and vascular inflammation using SUV, TBR, and a global parameter called the metabolic volumetric product (MVP). These approaches may be applied to assess vascular inflammation in various study samples of interest in a consistent fashion as we have shown in several prior publications.9,20,27,28

Quantification of Atherosclerotic Plaque Activity and Vascular Inflammation using [18-F] Fluorodeoxyglucose Positron Emission Tomography/Computed Tomography FDG-PET/CT

There is great need to identify atherosclerosis non-invasively, and here we demonstrate how FDG-PET/CT can be used to detect and quantify atherosclerotic plaque activity and vascular inflammation.

アテローム性動脈硬化プラーク活動と血管の炎症の定量化を用いて[18 F]フルオロ陽電子放射断層撮影/コンピュータ断層撮影（FDG-PET/CT）

Conventional non-invasive imaging modalities of atherosclerosis such as coronary artery calcium (CAC)1 and carotid intimal medial thickness (C-IMT)2 ...

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 ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely related to metabolism and energy expenditure, and its dysfunction is one important contributor for obesity and diabetes. Contrary to previous belief, recent PET/CT imaging studies indicated the BAT depots are still present in human adults. PET imaging clearly shows that BAT has considerably high uptake of 18F-FDG under certain conditions. In this video report, we demonstrate that Cerenkov luminescence imaging (CLI) with 18F-FDG can be used to optically image BAT in small animals. BAT activation is observed after intraperitoneal injection of norepinephrine (NE) and cold treatment, and depression of BAT is induced by long anesthesia. Using multiple-filter Cerenkov luminescence imaging, spectral unmixing and 3D imaging reconstruction are demonstrated. Our results suggest that CLI with 18F-FDG is a practical technique for imaging BAT in small animals, and this technique can be used as a cheap, fast, and alternative imaging tool for BAT research.

In this video report, we show the application of Cerenkov Luminescence Imaging (CLI) for interscapular brown adipose tissue in mice under activated and depressed conditions.

間褐色脂肪組織のチェレンコフ発光イメージング

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Localization, Identification, and Excision of Murine Adipose Depots

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 reached a staggering sum of $190.2 billion in direct medical costs alone. Obesity is a major risk factor for a wide host of diseases. Historically, little was known regarding adipose and its major and essential functions in the body. Brown and white adipose are the two main types of adipose but current literature has identified a new type of fat called brite or beige adipose. Research has shown that adipose depots have specific metabolic profiles and certain depots allow for a propensity for obesity and other related disorders. The goal of this protocol is to provide researchers the capacity to identify and excise adipose depots that will allow for the analysis of different factorial effects on adipose; as well as the beneficial or detrimental role adipose plays in disease and overall health. Isolation and excision of adipose depots allows investigators to look at gross morphological changes as well as histological changes. The adipose isolated can also be used for molecular studies to evaluate transcriptional and translational change or for in vitro experimentation to discover targets of interest and mechanisms of action. This technique is superior to other published techniques due to the design allowing for isolation of multiple depots with simplicity and minimal contamination.

Due to the drastic and negative connection between obesity and other comorbidities, research on the role adipose plays in disease and overall health is warranted. We present a protocol for the isolation and excision of adipose depots allowing for the study of adipose using in situ and in vitro methods.

マウス脂肪デポのローカライズ、識別、および切除

Obesity has increased dramatically in the last few decades and affects over one third of the adult US population. The economic effect of obesity in 2005 ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

移植のためのカプセル化サーモ前脂肪細胞の脂肪組織デポへ

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

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

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

顕微鏡を用いたひと脂肪組織血管機能評価

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

Characterization of Immune Cells in Human Adipose Tissue by Using Flow Cytometry

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the development of obesity-associated complications such as type 2 diabetes. To quantitatively and qualitatively investigate the immune cell subsets in human AT deposits, we have developed a flow cytometry approach. The stromal vascular fraction (SVF), containing the immune cells, is isolated from subcutaneous and visceral AT biopsies by collagenase digestion. Adipocytes are removed after centrifugation. The SVF cells are stained for multiple membrane-bound markers selected to differentiate between immune cell subsets and analyzed using flow cytometry. As a result of this approach, pro- and anti-inflammatory macrophage subsets, dendritic cells (DCs), B-cells, CD4+ and CD8+ T-cells, and NK cells can be detected and quantified. This method gives detailed information about immune cells in AT and the amount of each specific subset. Since there are numerous fluorescent antibodies available, our flow cytometry approach can be adjusted to measure various other cellular and intracellular markers of interest.

This article describes a method to analyze immune cell content of adipose tissue by isolation of immune cells from adipose tissue and subsequent analysis using flow cytometry.

フローサイトメトリーを用いたひと脂肪組織での免疫細胞のキャラクタリゼーション

Infiltration of immune cells in the subcutaneous and visceral adipose tissue (AT) deposits leads to a low-grade inflammation contributing to the ...

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 ...

Continuous Blood Sampling in Small Animal Positron Emission Tomography/Computed Tomography Enables the Measurement of the Arterial Input Function

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal blood time-activity concentration also known as arterial input function (AIF) is a key point, especially for the characterization of animal disease models and the introduction of newly developed radiotracers. The knowledge of radiotracer availability in the blood helps to interpret PET/CT-derived data of tissue activity. For this purpose, online blood sampling during the PET/CT imaging is advisable to measure the AIF. In contrast to manual blood sampling and image-derived approaches, continuous online blood sampling has several advantages. Besides the minimized blood loss, there is an improved resolution and a superior accuracy for the blood activity measurement. However, the major drawback of online blood sampling is the costly and time-consuming preparation to catheterize the femoral vessels of the animal. Here, we describe an easy and complete workflow for catheterization and continuous blood sampling during small animal PET/CT imaging and compared it to manual blood sampling and an image-derived approach. Using this highly-standardized workflow, the determination of the fluorodeoxyglucose ([18F]FDG) AIF is demonstrated. Further, this procedure can be applied to any radiotracer in combination with different animal models to create fundamental knowledge of tracer kinetic and model characteristics. This allows a more precise evaluation of the behavior of pharmaceuticals, both for diagnostic and therapeutic approaches in the preclinical research of oncological, neurodegenerative and myocardial diseases.

Here a protocol for continuous blood sampling during PET/CT imaging of rats to measure the arterial input function (AIF) is described. The catheterization, the calibration and setup of the system and the data analysis of the blood radioactivity are demonstrated. The generated data provide input parameters for subsequent bio-kinetic modeling.

小動物陽電子放出断層撮影/コンピュータ断層撮影における連続的な血液サンプリングにより、動脈入力機能の測定が可能

For quantitative analysis and bio-kinetic modeling of positron emission tomography/computed tomography (PET/CT) data, the determination of the temporal ...