博士カルバリョの仕事は女性の健康と母子保健と米国立衛生研究所（5K12 HD043452）の人間開発の国立研究所で研究局からの研究助成金によって支えられている。博士の不安は、I-フロー（レイクフォレスト、カリフォルニア州）からの供給（オンQ PainBuster術後痛みを軽減するシステム）と資金（生化学的アッセイ用）を受け取った。

1。研究プロトコル<ul><li>治験審査委員会の承認と書面によるインフォームドコンセントを取得します。 </li><li>研究実施のための適切な外科的集団を選択します。この方法論は、帝王切開の傷に適用されているが、ほとんどの手術の傷は適切でしょう。 </li><li>あなたの研究参加のための包含と除外の基準を考慮してください。手術創感染率の高い患者と手順は適切でないかもしれません。 </li><li>潜在的な交絡因子を避けるために、すべての患者のための麻酔および術後管理を標準化します。 </li></ul> 2。侵害受容性および炎症性メディエータ生化学コレクション<ul><li>閉鎖（他の組織層が考えられる）を巻か直前に皮下層に、ON-Q SilverSoaker（I-フロー、レイクフォレスト、カリフォルニア州）でON-Q PainBusterを挿入します。 ON-Q PainBusterの痛みを軽減するシステムが継続的に2ミリリットル/ hの速度で生理食塩水を提供します。キャリアは櫛することができますとinedまたはそのような局所麻酔薬および非ステロイド性抗炎症薬のような目的の薬に比べて。 </li><li>指定された時点で創傷滲出液の吸引を可能にするためにこのシステムに標準の三方活栓を組み込んでいる。連続注入、凝固からカテーテルを防止し、滲出液サンプルを生成するために、システムの信頼性を向上させます。滲出液の吸引が困難な場合（例約5％）、傷の上にそっと押す（ 例えば 、患者が座ったりが平らに横たわって）被写体の位置を変え、0.5〜1ミリリットル生理食塩水フラッシュを使用するか、撤退を検討カテーテル1〜2センチメートル。 </li><li>プロトコル（ 例えば 、1、4、6、24、帝王切開分娩48時間後）で指定された時点で、創傷滲出液の1ミリリットルを吸引し、撤退する。プロテイナーゼインヒビターの30μlを含むポリエチレンカップに滲出液を移す。我々は、我々の目的のために完全なプロテイナーゼ阻害剤（バイオサイエンスロシュ、パロアルト、CA）を使用しています。 </li><li&gt;治験実施計画書で指定された同じ時間間隔で、血液の10ミリリットルを収集します。我々は、リチウムヘパリンを含む採血管を使用していた、我々の施設で、これは緑で覆われたガラス管である。血液サンプルにプロテイナーゼ阻害剤の追加の300μlを添加する。 </li><li>コレクションの1時間以内に、10分間3,000 rpmで氷と遠心分離機にサンプルを置く。 </li><li> -20℃での標準的なマイクロ遠心チューブや店舗における血清および創傷清と場所を削除する</li></ul> 3。アッセイ分析<ul><li>一度すべての研究滲出および血清試料を収集し、保存、解凍し、同時にサンプルを分析しています。 </li><li>我々は、マルチプレックスイムノアッセイを用いてサイトカインやケモカインを測定した。多重免疫アッセイ技術は、私たちは小さい（50μl）をボリュームサンプルから多数の検体をアッセイすることができます。マルチプレックスは、ELISA法を用いて得られたものと同等の結果を生成します。我々は17プレックスバイオPlexTMシステム（バイオラッド、ハークを使用していた我々の分析のためにULES、カリフォルニア州）。このアッセイは、腫瘍壊死因子α（TNFα）、インターフェロンα（INFα）、インターロイキン1β（IL-1β）、インターロイキン2（IL-2）、インターロイキン4（IL-4）、インターロイキン-5（IL-5を測定することが可能である）、インターロイキン-6（IL-6）、インターロイキン7（IL-7）、インターロイキン-8（IL-8）、インターロイキン（IL-10）、インターロイキン12（IL-12）、インターロイキン13（IL-13）、インターロイキン17（IL-17）、顆粒球コロニー刺激因子（G-CSF）、顆粒球マクロファージコロニー刺激因子（GM-CSF）、単球走化性タンパク質-1（MCP-1）、マクロファージ炎症性タンパク質（MIP-1β）。神経成長因子は、Bio-Plexのアミンカップリングキットと17-プレックスのプレートを追加することで、NGF DY256抗体を用いて測定した。 </li><li>良い品質の分析を確実にするために、各測定が重複してなされるべきである。各分析対象成分の標準曲線は（0.20の濃度で、0.78、3.13、12.5、50、200、800、3200 pg / mlのプラスメーカーによって供給される基準の検体を使用して生成され生理食塩水はゼロ標準）。標準曲線は、各ランに含まれており、試料濃度は、Bio-Plexのマネージャーソフトウェアで計算されます。 </li><li>サブスタンスPおよびプロスタグランジンE2は非常に感度のELISAキットを用いて測定した。我々は、この分析のためにアッセイデザイン社（アナーバー、ミシガン州）が使用されています。メーカーの仕様によると、二重に分析を実行します。 </li></ul>

<ol>
	<li>Carvalho, B., Clark, D. J., Angst, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Local+and+Systemic+Release+of+Cytokines,+Nerve+Growth+Factor,+Prostaglandin+E2,+and+Substance+P+in+Incisional+Wounds+and+Serum+Following+Cesarean+Delivery.">Local and Systemic Release of Cytokines, Nerve Growth Factor, Prostaglandin E2, and Substance P in Incisional Wounds and Serum Following Cesarean Delivery.</a> J. Pain. 9, 650-657 (2008).</li><li>Carvalho, B., Clark, D. J., Yeomans, D. C., Angst, M. 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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+epidural+clonidine+on+perioperative+cytokine+response,+postoperative+pain,+and+bowel+function+in+patients+undergoing+colorectal+surgery.">The effect of epidural clonidine on perioperative cytokine response, postoperative pain, and bowel function in patients undergoing colorectal surgery.</a> Anesth. Analg. 99, 502-509 (2004).</li><li>Buvanendran, A., Kroin, J. S., Berger, R. A., Hallab, N. J., Saha, C., Negrescu, C., Moric, M., Caicedo, M. S., Tuman, K. 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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficacy+of+continuous+wound+catheters+delivering+local+anesthetic+for+postoperative+analgesia:+a+quantitative+and+qualitative+systematic+review+of+randomized+controlled+trials.">Efficacy of continuous wound catheters delivering local anesthetic for postoperative analgesia: a quantitative and qualitative systematic review of randomized controlled trials.</a> J. Am. Coll. Surg. 203, 914-932 (2006).</li><li>Eriksson, A. S., Sinclair, R., Cassuto, J., Thomsen, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Influence+of+lidocaine+on+leukocyte+function+in+the+surgical+wound.">Influence of lidocaine on leukocyte function in the surgical wound.</a> Anesthesiology. 77, 74-78 (1992).</li><li>Clark, J. D., Shi, X., Li, X., Qiao, Y., Liang, D., Angst, M. S., Yeomans, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Morphine+reduces+local+cytokine+expression+and+neutrophil+infiltration+after+incision.">Morphine reduces local cytokine expression and neutrophil infiltration after incision.</a> Mol. Pain. 3, 28 (2007).</li><li>Webb, S. T., Ghosh, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intra-articular+bupivacaine:+potentially+chondrotoxic.">Intra-articular bupivacaine: potentially chondrotoxic.</a> Br. J. Anaesth. 102, 439-441 (2009).</li><li>Scott, N. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+infiltration+for+surgery.">Wound infiltration for surgery.</a> Anaesthesia. 65, 67 (2010).</li></ol>

特別な利害関係は宣言されません。

上記のこの方法は、創傷滲出液生化学的炎症と侵害受容性メディエーターを検出するのに十分な感度であることが判明している。各種の時点でのサンプリングはまた、私たちは時間をかけてこれらのメディエーターの変化を観察することができました。1,2は、我々 は逐次外科創傷滲出液における炎症と侵害受容性メディエーターの放出プロファイルを測定する他のどの人間?...

外科的切開した後、常駐し、移行炎症細胞が痛みを開始および維持するために重要であるプロと抗炎症性サイトカイン、プロスタノイド、神経ペプチドとケモカインを放出する。3,4、サイト固有の生化学的マーカーを収集する能力を決定することが重要です血清および手術創内のレベル間の関係は、メディエーター放出と術後疼痛との間の複雑な関係を理解し​​、手術創生化学上の両方​​の全身および末梢薬剤投与の効果を評価できるようにする。研究は、生化学的メディエーターの全身測定に焦点を当てていると、サイト固有のリリースは考慮していません。サイトカインおよびケモカインの5,6血清測定値が局所手術創生化学的事象を反映するものとだけ外科的損傷へのグローバルな全身反応を示さないことがあります。この記事では、モミだったこの方法論の詳細とユニークな視覚的な説明を提供しています痛みのジャーナルで2008年に参照聖1は、このJoveの記事の目的は、メソッドのユニークで視覚的に強化された説明を提供することです。

collecting and measuring wound exudate biochemical mediators in surgical wounds

我々は、我々が手術創傷部位での炎症性および生化学的な侵害受容性メディエーターを収集し、測定することができますそれによって方法論を記述する。部位特異的な生化学的マーカーを収集すると、血清中および手術創内のレベル間の関係を理解するメディエーター放出、痛み、鎮痛剤の使用および関心のある他の転帰との関連付けを決定し、手術創に全身および末梢薬物投与の効果を評価することは重要である生化学。この手法は、脊椎麻酔による選択的帝王切開分娩を受けて健康な女性に適用されています。私たちは、患者の疼痛スコアと最大48時間後に帝王切開分娩のための鎮痛剤の消費量と同じ時間間隔で創傷滲出液および血清メディエーターを測定した。この方法論を使用して我々は、神経成長因子（NGF）、プロスタグランジンE2（PG-E2）のサブスタンスPは、IL-1β、IL-2、IL-4、IL-6、IL-7など、様々な生化学メディエーターを検出することができましたIL-8、IL-10、IL-12、IL-13、IL-17、TNFα、INFγ、G-CSF、GM-CSF、MCP-1およびMIP-1β。この人間の手術創バイオアッセイを適用した研究は、創傷および血清サイトカイン濃度またはその時間の放出プロファイルの間には相関関係は認められません（J疼痛2008;。9（7）:650-7）1我々はまた、識別する手法の有用性を文書化した。薬剤媒介創傷サイトカイン量の変化（Anesth Analg 2010; 111:1452-9）。2

この方法論を使用して、我々は、創傷滲出液および血漿中のIL-5の例外を除いて上記のすべての生化学メディエーターを検出することができました。1,2いくつかの追加のサイトカインが（TNFα、IL-4、IL-17、、血漿中では検出されなかったINFγ 、GM-CSF）。報告された研究で測定されたサイトカインおよびケモカインの1時間コースでは、24時間の観察期間持続した最初の数時間以内...

Watch this Scientific Journal Video about Collecting And Measuring Wound Exudate Biochemical Mediators In Surgical Wounds at JoVE.com

Collecting And Measuring Wound Exudate Biochemical Mediators In Surgical Wounds

この記事では、帝王切開後に手術創傷部位での炎症性および生化学的な侵害受容性メディエーターを収集し、測定するための方法論の詳細と視覚的な説明を提供しています。この人間のバイオアッセイは、創傷および血清サイトカイン濃度および創傷サイトカイン、ケモカインおよび神経ペプチドにおける薬剤性の変化との相関関係を決定するために使用されています。

手術創内に創傷滲出液生化学メディエーターの収集と測定

We describe a methodology by which we are able to collect and measure biochemical inflammatory and nociceptive mediators at the surgical wound site. ...

collecting-measuring-wound-exudate-biochemical-mediators-surgical

Research

JoVE Journal

Medicine

11.9K ビュー.  Stanford University School of Medicine. この記事では、帝王切開後に手術創傷部位での炎症性および生化学的な侵害受容性メディエーターを収集し、測定するための方法論の詳細と視覚的な説明を提供しています。この人間のバイオアッセイは、創傷および血清サイトカイン濃度および創傷サイトカイン、ケモカインおよび神経ペプチドにおける薬剤性の変化との相関関係を決定するために使?...

ビデオ: Collecting And Measuring Wound Exudate Biochemical Mediators In Surgical Wounds

Biochemical Measurement of Neonatal Hypoxia

Neonatal hypoxia ischemia is characterized by inadequate blood perfusion of a tissue or a systemic lack of oxygen. This condition is thought to cause/exacerbate well documented neonatal disorders including neurological impairment 1-3. Decreased adenosine triphosphate production occurs due to a lack of oxidative phosphorylation. To compensate for this energy deprived state molecules containing high energy phosphate bonds are degraded 2. This leads to increased levels of adenosine which is subsequently degraded to inosine, hypoxanthine, xanthine, and finally to uric acid. The final two steps in this degradation process are performed by xanthine oxidoreductase. This enzyme exists in the form of xanthine dehydrogenase under normoxic conditions but is converted to xanthine oxidase (XO) under hypoxia-reperfusion circumstances 4, 5. Unlike xanthine dehydrogenase, XO generates hydrogen peroxide as a byproduct of purine degradation 4, 6. This hydrogen peroxide in combination with other reactive oxygen species (ROS) produced during hypoxia, oxidizes uric acid to form allantoin and reacts with lipid membranes to generate malondialdehyde (MDA) 7-9. Most mammals, humans exempted, possess the enzyme uricase, which converts uric acid to allantoin. In humans, however, allantoin can only be formed by ROS-mediated oxidation of uric acid. Because of this, allantoin is considered to be a marker of oxidative stress in humans, but not in the mammals that have uricase.
We describe methods employing high pressure liquid chromatography (HPLC) and gas chromatography mass spectrometry (GCMS) to measure biochemical markers of neonatal hypoxia ischemia. Human blood is used for most tests. Animal blood may also be used while recognizing the potential for uricase-generated allantoin. Purine metabolites were linked to hypoxia as early as 1963 and the reliability of hypoxanthine, xanthine, and uric acid as biochemical indicators of neonatal hypoxia was validated by several investigators 10-13. The HPLC method used for the quantification of purine compounds is fast, reliable, and reproducible. The GC/MS method used for the quantification of allantoin, a relatively new marker of oxidative stress, was adapted from Gruber et al 7. This method avoids certain artifacts and requires low volumes of sample. Methods used for synthesis of MMDA were described elsewhere 14, 15. GC/MS based quantification of MDA was adapted from Paroni et al. and Cighetti et al. 16, 17. Xanthine oxidase activity was measured by HPLC by quantifying the conversion of pterin to isoxanthopterin 18. This approach proved to be sufficiently sensitive and reproducible.

A method is described to measure biochemical markers of neonatal hypoxia-ischemia. The approach utilizes high pressure liquid chromatography (HPLC) and Gas Chromatography Mass Spectrometry (GC/MS).

新生児低酸素症の生化学的測定

Neonatal hypoxia ischemia is characterized by inadequate blood perfusion of a tissue or a systemic lack of oxygen. This condition is thought to ...

医学

Mouse Model of Intraluminal MCAO: Cerebral Infarct Evaluation by Cresyl Violet Staining

Stroke is the third cause of mortality and the leading cause of disability in the World. Ischemic stroke accounts for approximately 80% of all strokes. However, the thrombolytic tissue plasminogen activator (tPA) is the only treatment of acute ischemic stroke that exists. This led researchers to develop several ischemic stroke models in a variety of species. Two major types of rodent models have been developed: models of global cerebral ischemia or focal cerebral ischemia. To mimic ischemic stroke in patients, in whom approximately 80% thrombotic or embolic strokes occur in the territory of the middle cerebral artery (MCA), the intraluminal middle cerebral artery occlusion (MCAO) model is quite relevant for stroke studies. This model was first developed in rats by Koizumi et al. in 1986 1. Because of the ease of genetic manipulation in mice, these models have also been developed in this species 2-3.
	Herein, we present the transient MCA occlusion procedure in C57/Bl6 mice. Previous studies have reported that physical properties of the occluder such as tip diameter, length, shape, and flexibility are critical for the reproducibility of the infarct volume 4. Herein, a commercial silicon coated monofilaments (Doccol Corporation) have been used. Another great advantage is that this monofilament reduces the risk to induce subarachnoid hemorrhages. Using the Zeiss stereo-microscope Stemi 2000, the silicon coated monofilament was introduced into the internal carotid artery (ICA) via a cut in the external carotid artery (ECA) until the monofilament occludes the base of the MCA. Blood flow was restored 1 hour later by removal of the monofilament to mimic the restoration of blood flow after lysis of a thromboembolic clot in humans. The extent of cerebral infarct may be evaluated first by a neurologic score and by the measurement of the infarct volume. Ischemic mice were thus analyzed for their neurologic score at different post-reperfusion times. To evaluate the infarct volume, staining with 2,3,5-triphenyltetrazolium chloride (TTC) was usually performed. Herein, we used cresyl violet staining since it offers the opportunity to test many critical markers by immunohistochemistry. In this video, we report the MCAO procedure; neurological scores and the evaluation of the infarct volume by cresyl violet staining.

The intraluminal middle cerebral occlusion model in mice is herein presented. The extent of cerebral infarct is evaluated by a neurologic score and cresyl violet staining, an alternative staining to TTC, offering the great advantage to test in parallel many interest markers.

腔内MCAOのマウスモデル：クレシルバイオレット染色による脳梗塞の評価

Stroke is the third  cause of mortality and the leading cause of disability in the World. Ischemic  stroke accounts for approximately 80% of all strokes. ...

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research that are easy-to-collect, minimally invasive, and relatively inexpensive biologic markers of stress. This article we present the steps for collection and analysis of two salivary assays in research with frail, community residing older adults-salivary cortisol and salivary alpha amylase. The field of salivary bioscience is rapidly advancing and the purpose of this presentation is to provide an update on the developments for investigators interested in integrating these measures into research on aging. Strategies are presented for instructing family caregivers in collecting saliva in the home, and for conducting laboratory analyses of salivary analytes that have demonstrated feasibility, high compliance, and yield quality specimens. The protocol for sample collection includes: (1) consistent use of collection materials; (2) standardized methods that promote adherence and minimize subject burden; and (3) procedures for controlling certain confounding agents. We also provide strategies for laboratory analyses include: (1) saliva handling and processing; (2) salivary cortisol and salivary alpha amylase assay procedures; and (3) analytic considerations.

We demonstrate: (1) procedures for collection of salivary samples in cognitive impaired older adults by family caregivers in the home setting, (2) procedures for measuring stress activity via salivary cortisol and alpha amylase, and (3) representative profiles. Protocols that allow researchers to study stress-linked processes advance our understanding of biological sensitivity and susceptibility.

唾液を収集し、家族介護者を経由して高齢者を居住する虚弱コミュニティ唾液コルチゾールおよびα-アミラーゼの測定

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease and injury regional estimates, World Health Organization, 2004). Surgeries that often compromise the quality of life are required to correct heart defects, reminding us of the importance of finding the causes of CHD. Mutant mouse models and live imaging technology have become essential tools to study the etiology of this disease. Although advanced methods allow live imaging of abnormal hearts in embryos, the physiological and hemodynamic states of the latter are often compromised due to surgical and/or lengthy procedures. Noninvasive ultrasound imaging, however, can be used without surgically exposing the embryos, thereby maintaining their physiology. Herein, we use simple M-mode ultrasound to assess heart rates of embryos at E18.5 in utero. The detection of abnormal heart rates is indeed a good indicator of dysfunction of the heart and thus constitutes a first step in the identification of developmental defects that may lead to heart failure.

In utero Measurement of Heart Rate in Mouse by Noninvasive M-mode Echocardiography

Ultra-high frequency ultrasound is a powerful live imaging tool to examine cardiac abnormalities in small animals. Its noninvasiveness allows maintaining the physiologic state of embryos. Herein, we show the use of M-mode ultrasound to measure the heart rates of embryos at E18.5 in utero.

非侵襲Mモード心エコー検査によるマウスでの心拍数の子宮内測定で

Congenital heart disease (CHD) is the most frequent noninfectious cause of death at birth. The incidence of CHD ranges from 4 to 50/1,000 births (Disease ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

腫瘍関連血管系を使用しての生体顕微鏡が背高度な蛍光トランスジェニック マウスの皮下脂肪のウィンドウの部屋

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an otherwise dynamic system, and successive phases are easily overlooked or misinterpreted. Live-cell imaging and time-lapse microscopy, in which living cells can be followed for hours or even days in a more or less continuous fashion, are therefore very informative. The protocol described here allows for the investigation of the fate of chemotherapeutic nanoparticles after the delivery of doxorubicin (dox) in living cells. Dox is an intercalating agent that must be released from its nanocarrier to become biologically active. In spite of its clinical registration for more than two decades, its uptake, breakdown, and drug release are still not fully understood. This article explores the hypothesis that lipid-based nanoparticles are taken up by the tumor cells and are slowly degraded. Released dox is then translocated to the nucleus. To prevent fixation artifacts, live-cell imaging and time-lapse microscopy, described in this experimental procedure, can be applied.

Using In Vitro Live-cell Imaging to Explore Chemotherapeutics Delivered by Lipid-based Nanoparticles

Live-cell imaging is a powerful tool to visualize dynamic processes. The examination of fixed cells provides only static pictures, which can lead to misinterpretation and confusion about the process. This work presents a method to study uptake, drug release, and intracellular localization of liposomal nanoparticles in living cells.

体外配信生細胞イメージング化学療法を探求する脂質ナノ粒子によってを使用してください。

Conventional imaging techniques can provide detailed information about cellular processes. However, this information is based on static images in an ...

Collecting Hair Samples for Hair Cortisol Analysis in African Americans

The hormone cortisol is typically assessed in saliva, serum, or urine samples. More recently, cortisol has been successfully extracted from hair, including humans. The advantage of hair cortisol concentration is that it reflects a retrospective representation of hypothalamic-pituitary-adrenal (HPA) axis function over time, much like hemoglobin A1C represents glycemic control. However, obtaining hair samples can be challenging, due to the cultural beliefs and hair care practices of minority participants. For example, African Americans may be reluctant to provide samples. Additionally, few researchers are trained to collect hair samples from African Americans. The purpose of this paper is to present a culturally informed protocol to help researchers obtain hair samples from African Americans. To illustrate the representative results of this protocol implementation, de-identified data from African Americans that participated in a community-based study on chronic stress are provided. Hair practice preferences are assessed. The participants are made comfortable by showing pictures of hair samples prior to cutting their hair. The single strain twist and gently pull method is used to collect approximately 30 - 50 strands of hair from the posterior vertex region of the scalp. This protocol will significantly improve collection of hair samples from African Americans.

Hair cortisol concentration analysis provides an alternative to traditional measures of cortisol; however, to collect hair samples from African Americans, scientists need to be culturally informed and competent. The purpose of this protocol is to demonstrate a culturally informed technique to collect hair samples for cortisol analysis from African Americans.

髪コルチゾールにおけるアフリカ系アメリカ人の毛のサンプルの収集

The hormone cortisol is typically assessed in saliva, serum, or urine samples. More recently, cortisol has been successfully extracted from hair, ...

Protocol to Create Chronic Wounds in Diabetic Mice

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ~6.5M people and cost ~$40B/year in the US alone. Although a significant effort has been invested in understanding how chronic wounds develop in humans, fundamental questions remain unanswered. Recently, we developed a novel mouse model for diabetic chronic wounds that have many characteristics of human chronic wounds. Using db/db-/- mice, we can generate chronic wounds by inducing high levels of oxidative stress (OS) in the wound tissue immediately after wounding, using a one-time treatment with inhibitors specific to the antioxidant enzymes catalase and glutathione peroxidase. These wounds have high levels of OS, develop biofilm naturally, become fully chronic within 20 days after treatment and can remain open more for more than 60 days. This novel model has many features of diabetic chronic wounds in humans and therefore can contribute significantly to advancing fundamental understanding of how wounds become chronic. This is a major breakthrough because chronic wounds in humans cause significant pain and distress to patients and result in amputation if unresolved. Moreover, these wounds are very expensive and time-consuming to treat, and lead to significant loss of personal income to patients. Advancements in this field of study through the use of our chronic wound model can significantly improve health care for millions who suffer under this debilitating condition. In this protocol, we describe in great detail the procedure to cause acute wounds to become chronic, which has not been done before.

Chronic wounds are developed from acute wounds on a diabetic mouse model by inducing high levels of oxidative stress after a full-thickness cutaneous wound. The wound is treated with inhibitors for catalase and glutathione peroxidase, resulting in impaired healing and biofilm development by bacteria present in the skin microbiome.

糖尿病マウスの慢性創傷を作成するためのプロトコル

Chronic wounds develop as a result of defective regulation in one or more complex cellular and molecular processes involved in proper healing. They impact ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

角膜上皮摩耗眼はりのモデルとして角膜創傷治癒

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

Targeting Gray Rami Communicantes in Selective Chemical Lumbar Sympathectomy

Chemical lumbar sympathectomy (CLS) is a commonly used, minimally invasive procedure for the treatment of conditions including ischemic diseases of the lower extremities, hyperhidrosis, etc. It is commonly practiced to position the puncture needle tip in front of the anterior fascia of the psoas major muscle and inject the inactivating agent around the sympathetic trunk, which is defined as conventional CLS. Although relatively rare, ureteropelvic damage is the most frequently reported complication of conventional CLS and can cause serious harm to patients. We found that injecting the inactivating agent behind the anterior fascia, which only targets gray rami communicantes, helped achieve therapeutic efficacy in&#160;vasodilation, sweat reduction, and pain relief comparable to conventional CLS, and serious complications were largely reduced. We define this procedure as selective CLS. Here, we present a protocol of selective CLS. The precise needle tract and accurate evaluation of the spreading of the contrast agent are critical to ensure that the drug is injected behind the anterior fascia of the psoas major muscle. The needle tip is at approximately one-third the dividing line of the vertebral body in the lateral view of a lumbar X-ray. The contrast is mainly confined around the needle tip and spreads outward and downward along the psoas muscle fibers. In this way, the anterior fascia provides a natural barrier for the ureteropelvic area, and the psoas major muscle provides a natural barrier for the lumbar nerve root. There are several highlights of this article, including 1) a detailed description of the selective CLS procedures, 2) an explanation of the anatomical basis for the implementation of selective CLS, and 3) an explanation of the differences between selective and conventional CLS.

We present a protocol for selective chemical lumbar sympathectomy (CLS), which inactivates only gray rami communicantes and not the sympathetic trunk. Selective CLS can help achieve therapeutic efficacy in vasodilation, sweat reduction, and pain relief that are comparable to conventional CLS, and serious complications, particularly ureteropelvic damages, can be reduced.

ターゲット灰白交通枝選択的化学腰椎交感神経切除術で

Chemical lumbar sympathectomy (CLS) is a commonly used, minimally invasive procedure for the treatment of conditions including ischemic diseases of the ...