The authors acknowledge the Carefusion Foundation for support via a clinical excellence grant of this work titled &#8220;Prospective evaluation of procedural povidone iodine rectal preparation to reduce infectious complications following ultrasound guided prostate needle biopsy.&#8221;

この患者に関連する研究は、ヒトの研究のための施設内倫理委員会（IRB）により承認されました。 1.患者の準備と配置<ol><li>手順スイートに患者を持参し、膝が胸に屈曲して、その右側または左側に配置します。 </li><li>患者、外科的手順、および表示が見直され、確認されていることにより、外科的タイムアウトを実行します。 </li><li>示されたときは、麻酔サービスを介して静脈内鎮静を管理します。 </li></ol> 2.直腸指診（DRE）を実行<ol><li>両手にサイズ適切な手袋を置きます。 </li><li>直腸ボールトに潤滑した後、手袋をはめた指を挿入して、直腸超音波検査中の着目を必要としている可能性のある不審な分野のために感じるように前立腺を調べる。10 </li></ol>ポビドンヨード直腸の調製3.作成（PIRP） <ol><li> 10％の15ミリリットルを注ぎます無菌流域に市販ポビドンヨード液。 </li><li>この盆地に市販の1％リドカインゲルの5ミリリットルを追加します。 </li><li>ポビドンヨードスラリーを作成するために、15秒間の内容を混ぜます。 </li></ol> PIRP 4.プライマリ管理<ol><li>文化に滅菌綿棒直腸直腸ボールトを使用してください。 </li><li> 、4センチメートルX 4センチメートル滅菌ガーゼを開きポビドンヨードスラリーに浸し、その後、直腸ボールトに挿入します。ガーゼのわずかな尾が肛門から出てくる残されています。 </li><li>それは除去し、廃棄された後、2分間の代わりにガーゼを残します。 </li></ol> PIRP 5.セカンダリ管理<ol><li> PIRP溶液と肛門周囲の領域を描画するために使い捨て婦人科の綿棒を使用してください。 </li><li>肛門から3センチメートルの距離に直腸ボールトにこの婦人科綿棒を挿入します。 </li><li>さらにPIRPに直腸粘膜の露出を最大化するために、直腸VAUを綿棒同様にして、LTの少なくとも3倍。 </li><li>ポビドンヨード溶液を2のために乾燥することができます- 。前の標準的な経直腸超音波検査とその後の生検を進めるに3分11 PIRPの殺菌効果が露出粘膜上で乾燥するためのソリューションを必要とし、これは不可欠です。 </li><li>文化に滅菌綿棒直腸直腸ボールトを使用してください。 </li></ol> 6.経直腸超音波検査と生検<ol><li>直腸を経由して超音波プローブを挿入します。 </li><li>すべての泌尿器慣行と一致し、標準的な方法で前立腺生検およびその後の測定を行う。11,12 </li></ol>

<ol>
	<li>Nam, R. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increasing+hospital+admission+rates+for+complications+after+transrectal+ultrasound+guided+prostate+biopsy.">Increasing hospital admission rates for complications after transrectal ultrasound guided prostate biopsy.</a> J Urol. 189, S12-S17 (2013).</li><li>Feliciano, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+incidence+of+fluoroquinolone+resistant+infections+after+prostate+biopsy+are+fluoroquinolones+still+effective+prophylaxis.">The incidence of fluoroquinolone resistant infections after prostate biopsy are fluoroquinolones still effective prophylaxis.</a> J Urol. 179 (3), 952-955 (2008).</li><li>Kehinde, O., Al-Maghrebi, M., Sheikh, M., Anim, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combined+ciprofloxacin+and+amikacin+prophylaxis+in+the+prevention+of+septicemia+after+transrectal+ultrasound+guided+biopsy+of+the+prostate.">Combined ciprofloxacin and amikacin prophylaxis in the prevention of septicemia after transrectal ultrasound guided biopsy of the prostate.</a> J Urol. 189 (3), 911-915 (2013).</li><li>Taylor, A. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeted+antimicrobial+prophylaxis+using+rectal+swab+cultures+in+men+undergoing+transrectal+ultrasound+guided+prostate+biopsy+is+associated+with+reduced+incidence+of+postoperative+infectious+complications+and+cost+of+care.">Targeted antimicrobial prophylaxis using rectal swab cultures in men undergoing transrectal ultrasound guided prostate biopsy is associated with reduced incidence of postoperative infectious complications and cost of care.</a> J Urol. 187 (4), 1275-1279 (2012).</li><li>Gyorfi, J. R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peri+procedural+povidone+iodine+rectal+preparation+reduces+microorganism+counts+and+infectious+complications+following+ultrasound-guided+needle+biopsy+of+the+prostate.">Peri procedural povidone iodine rectal preparation reduces microorganism counts and infectious complications following ultrasound-guided needle biopsy of the prostate.</a> World J Urol. 32 (4), 905-909 (2014).</li><li>Park, D. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Control+of+infective+complications+of+transrectal+prostate+biopsy.">Control of infective complications of transrectal prostate biopsy.</a> Surg Infect Larchmt. 15 (4), 431-436 (2014).</li><li>Park, D. S., Oh, J. J., Lee, J. H., Jang, W. K., Hong, Y. K., Hong, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Simple+use+of+the+suppository+type+povidone+iodine+can+prevent+infectious+complications+in+transrectal+ultrasound+guided+prostate+biopsy.">Simple use of the suppository type povidone iodine can prevent infectious complications in transrectal ultrasound guided prostate biopsy.</a> Adv Urol. , (2009).</li><li>AbuGhosh, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+prospective+randomized+trial+of+povidone+iodine+prophylactic+cleansing+of+the+rectum+before+transrectal+ultrasound+guided+prostate+biopsy.">A prospective randomized trial of povidone iodine prophylactic cleansing of the rectum before transrectal ultrasound guided prostate biopsy.</a> J Urol. 189 (4), 1326-1331 (2013).</li><li>Gerber, G. S., Brendler, C. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+the+Urological+Patient:+History,+Physical+Exam,+and+Urinalysis.">Evaluation of the Urological Patient: History, Physical Exam, and Urinalysis.</a> Campbell Walsh Urology. , (2015).</li><li>Hodge, K. K., McNeal, J. E., Terris, M. K., Stamey, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Random+systematic+versus+directed+ultrasound+guided+transrectal+core+biopsies+of+the+prostate.">Random systematic versus directed ultrasound guided transrectal core biopsies of the prostate.</a> J Urol. 142 (1), 71-74 (1989).</li><li>Donoghue, P. M., McSweeney, S. E., Jhaveri, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genitourinary+imaging:+current+and+emerging+applications.">Genitourinary imaging: current and emerging applications.</a> J Postgrad Med. 56 (2), 131-139 (2010).</li></ol>

The authors have nothing to disclose.

TRUS PNB次感染は非常に血管前立腺に直腸ボールト細菌の転位に続発しています。この原理に基づいて、我々は、前立腺針生検の前に微生物のコロニー数を減らすために防腐剤ポビドンヨードとしてトピックを使用する簡単な方法を探求してきました。私たちの経験は、生検の時にこのPIRPレジメンを用いた0.6％（P = 0.02）の4.3％から減少を強調しています。さらに、我々は直腸ボールト微生物?...

前立腺癌は男性で最も一般的な癌および癌関連死亡原因の第2位です。 2014年に、約233000例はほとんど3万男性はこの悪性腫瘍に屈すると、米国で診 ​​断されます。1、前立腺特異抗原（PSA）血液検査と直腸指診（DRE）は前立腺癌スクリーニングにおいて重要な役割を果たしているが、経直腸超音波ガイド付き前立腺針生検（TRUS PNB）が組織学的組織診断を得るための最も一般的な手段です。 TRUS PNBは、患者の直腸内に超音波プローブを挿入して、一般的にちょうど前方に位置して前立腺の周辺ゾーンの12月14日生検を取得することを含みます。各針生検は非常に血管前立腺に直腸壁を介して18 G針の通過を必要とします。そのため、出血や細菌転移に起因する感染の理論上のリスクは以下の手順が存在します。なしtheless、数理計算上の合併症率後の手順では、歴史的に低いままでいます。 前TRUS PNBに抗生物質予防法は、日常的に最も頻繁に使用される薬剤であるキノロン系の抗生物質を処方されています。適切な予防にもかかわらず、現代の研究では、生検後の感染性合併症の増加が関与。2はまた、研究では、キノロン耐性菌の出現、特にEに TRUS PNBを以下の感染性合併症でこの上昇を起因しています大腸菌 。3 キノロン耐性のこの上昇率は、TRUS PNB手順の代替予防レジメンの必要性を示唆しています。この目的を達成するために、いくつかの異なった道は調査中です。一つのアプローチは、経口キノロンと組み合わせて、生検時の静脈内または筋肉内の抗生物質の投与を含む。4この戦略はTRUS PNB以下敗血症事象を減少させることができるが、この方法の主な制限は、さらに、この抗生物質ファミリーに耐性菌を開発するための可能性です。別の最近調査方法は、5。キノロン耐性直腸フローラが定着患者をスクリーニングする前生検直腸スワブの使用を組み込むような生物が検出される場合、「標的」抗生物質レジメンを同定耐性細菌の感度プロファイルに基づいて使用することができます菌株。方法論はエレガントであるが、このような綿棒を取得する実際のプロセスは、選択的にキノロン選択培地で培養し、その後抗生物質を調整することは、多くの臨床実践に欠けすることができる臨床および検査インフラストラクチャが必要です。 生検の前に直腸ボールト微生物のコロニー数を減少させるために防腐話題の投与は、TRUS PNB感染を制限するための別の戦略を提示することができます。ポビドンiodoneは安価で、容易に入手可能な薬剤であります手術部位に適用されたときに減少した細菌数に文書化されました。両方の大腸や婦人科手術におけるアプリケーションはよく知られています。したがって、直腸製剤としてpovidineヨウ素の使用は、生検の前に追加の準備を必要とせずに縮小TRUS PNB感染症に、生産可能シンプル、かつ費用効果的な方法を提示します。泌尿器科文学に関しては、以前の仕事はTRUS PNB以下の臨床感染症の減少を指摘している。6-9この動画はPIRPの技術は、臨床実践に統合するために使いやすさとシンプルさを強調するために示しています。 TRUS PNBを受けたすべての患者は、最初は前立腺癌スクリーニングのための紹介や初期評価のための専門の泌尿器科クリニックで見られました。この評価は、患者の血清前立腺特異抗原（PSA）および直腸指診（DRE）の性能の評価を伴います。 PSAまたはDREのいずれかの異常をの推薦を促しましたTRUS PNB。 TRUS PNB手順が明らかにした感染症、出血、および尿閉を含むすべての患者と関連するリスクを検討しました。すべての患者は、生検の前にキノロン系の薬（ すなわち、シプロフロキサシン）、またはトリメトプリム-スルファメトキサゾール（バクトリム）のいずれかの経口抗生物質の予防の三日間を受け取りました。

<table border="0" cellpadding="0" cellspacing="0">
	<tbody>
		<tr>
			<td>Povidone Iodine Solution, 10% Topical Microbicide Antiseptic, 8 fl oz.</td>
			<td>Equate</td>
			<td>4067979</td>
			<td></td>
		</tr>
		<tr>
			<td>4x4 sterile gauze pads, white</td>
			<td>Dukal Corporation</td>
			<td>DKL1412</td>
			<td></td>
		</tr>
		<tr>
			<td>disposable gynecologic swab&#160;</td>
			<td>numerous vendors</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1% lidocaine gel&#160;</td>
			<td>numerous vendors</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

povidone iodine rectal preparation at time of prostate needle biopsy is a simple and reproducible means to reduce risk of procedural infection

シングル機関と集団ベースの研究では、経直腸超音波ガイド付き前立腺針生検（TRUS PNB）以下の感染性合併症が増加していることを強調しています。このような感染症は、直腸ボールトにコロニーを形成し、生検手順の間に、血流内に移行されているキノロン耐性微生物の大部分が起因しています。生検時のポビドンヨード直腸製剤（PIRP）は、TRUS PNBを以下の直腸微生物のコロニー数、したがって、結果として感染症を軽減するための簡単​​な、再現性のある方法です。 すべての患者は、生検の前に経口抗生物質治療の3日間投与されます。 PIRP技術は当初、TRUS PNBのための標準的な方法で患者を配置することを含みます。直腸指診後に、市販のポビドンヨードの10％溶液15mlのスラリーを作成するために、1％リドカインゼリーの5 mlを混合します。 4センチメートルX 4センチメートル滅菌ガーゼは、SOAですこのスラリーにヒツジシラミバエし、それが除去された後に2分間、直腸円蓋に挿入されます。その後、使い捨ての綿の婦人科用綿棒は、肛門周囲と肛門は3センチの距離に直腸ボールトの両方をペイントするために使用されます。前の標準的な経直腸超音波検査とその後の生検を進めるに3分 - ポビドンヨード溶液を次に2のために乾燥させます。 このPIRP技術は、4.3％から0.6％（P = 0.02）への生検後の感染症の関連減少と2012年の3月から当院で実際にありました。この予防計画の主な利点は感染を減らすために容易に入手可能で、安価な薬の使用と、そのシンプルさと再現性です。さらに、この技術は、多剤耐性生物の可能性をさらに伝播に追加抗生物質の全身投与に患者をさらす避けることができます。 TRUS PNBでPIRPの使用は、しかし、ヨウ素または貝A患者には適用されませんllergies。

このPIRP技術は、165人の患者がこの品質改善策に在籍されていたと（現在まで）2012年1月以来実践してきました。この時点では、ただ一人の患者（0.6％）は全身性敗血症の形で感染性合併症を経験しています。この患者は、多剤耐性大腸菌が定着しました大腸菌菌株。この患者にPIRPの投与後に直腸スワブ、直腸ボールトがまだ2.1×10 5 CFU / mlの細菌が、それによってPIRPが十...

Watch this Scientific Journal Video about Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection at JoVE.com

Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection

Here, we present a protocol for use at the time of transrectal ultrasound guided prostate needle biopsy (TRUS PNB) that is a simple and cost-effective means to reduce infections following the procedure.

前立腺針生検の時のポビドンヨード直腸準備手続き感染のリスクを軽減するためにシンプルで再現性のある手段であります

Single institution and population-based studies highlight that infectious complications following transrectal ultrasound guided prostate needle biopsy ...

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Research

JoVE Journal

Medicine

31.8K ビュー.  Penn State Milton S. Hershey Medical Center. Here, we present a protocol for use at the time of transrectal ultrasound guided prostate needle biopsy (TRUS PNB) that is a simple and cost-effective means to reduce infections following the procedure.

ビデオ: Povidone Iodine Rectal Preparation at Time of Prostate Needle Biopsy is a Simple and Reproducible Means to Reduce Risk of Procedural Infection

Evaluation of Nanoparticle Uptake in Tumors in Real Time Using Intravital Imaging

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle uptake in tumors at the microscopic level1,2,3, highlighting the utility of a suitable xenograft model in which to perform detailed uptake analyses. Here, we use high-resolution intravital imaging to evaluate nanoparticle uptake in human tumor xenografts in a modified, shell-less chicken embryo model. The chicken embryo model is particularly well-suited for these in vivo analyses because it supports the growth of human tumors, is relatively inexpensive and does not require anesthetization or surgery 4,5. Tumor cells form fully vascularized xenografts within 7 days when implanted into the chorioallantoic membrane (CAM) 6. The resulting tumors are visualized by non-invasive real-time, high-resolution imaging that can be maintained for up to 72 hours with little impact on either the host or tumor systems. Nanoparticles with a wide range of sizes and formulations administered distal to the tumor can be visualized and quantified as they flow through the bloodstream, extravasate from leaky tumor vasculature, and accumulate at the tumor site. We describe here the analysis of nanoparticles derived from Cowpea mosaic virus (CPMV) decorated with near-infrared fluorescent dyes and/or polyethylene glycol polymers (PEG) 7, 8, 9,10,11. Upon intravenous administration, these viral nanoparticles are rapidly internalized by endothelial cells, resulting in global labeling of the vasculature both outside and within the tumor7,12. PEGylation of the viral nanoparticles increases their plasma half-life, extends their time in the circulation, and ultimately enhances their accumulation in tumors via the enhanced permeability and retention (EPR) effect 7, 10,11. The rate and extent of accumulation of nanoparticles in a tumor is measured over time using image analysis software. This technique provides a method to both visualize and quantify nanoparticle dynamics in human tumors.

We present a novel approach to quantify nanoparticle localization in the vasculature of human xenografted tumors using dynamic, real-time intravital imaging in an avian embryo model.

生体内イメージングを使ってリアルタイムで腫瘍におけるナノ粒子の取り込みの評価

Current technologies for tumor imaging, such as ultrasound, MRI, PET and CT, are unable to yield high-resolution images for the assessment of nanoparticle ...

医学

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR (qPCR)

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process including development1, apoptosis2, and cell cycle regulation3. MiRNAs are estimated to regulate the expression of 30% to 90% of human genes4 by binding to their target messenger RNAs (mRNAs)5. Widespread dysregulation of miRNAs has been reported in various diseases and cancer subtypes6. Due to their prevalence and unique structure, these small molecules are likely to be the next generation of biomarkers, therapeutic agents and/or targets.
Methods used to investigate miRNA expression include SYBR green I dye- based as well as Taqman-probe based qPCR. If miRNAs are to be effectively used in the clinical setting, it is imperative that their detection in fresh and/or archived clinical samples be accurate, reproducible, and specific. qPCR has been widely used for validating expression of miRNAs in whole genome analyses such as microarray studies7. The samples used in this protocol were from patients who underwent radical prostatectomy for clinically localized prostate cancer; however other tissues and cell lines can be substituted in. Prostate specimens were snap-frozen in liquid nitrogen after resection. Clinical variables and follow-up information for each patient were collected for subsequent analysis8.
Quantification of miRNA levels in prostate tumor samples. The main steps in qPCR analysis of tumors are: Total RNA extraction, cDNA synthesis, and detection of qPCR products using miRNA-specific primers. Total RNA, which includes mRNA, miRNA, and other small RNAs were extracted from specimens using TRIzol reagent. Qiagen's miScript System was used to synthesize cDNA and perform qPCR (Figure 1). Endogenous miRNAs are not polyadenylated, therefore during the reverse transcription process, a poly(A) polymerase polyadenylates the miRNA. The miRNA is used as a template to synthesize cDNA using oligo-dT and Reverse Transcriptase. A universal tag sequence on the 5' end of oligo-dT primers facilitates the amplification of cDNA in the PCR step. PCR product amplification is detected by the level of fluorescence emitted by SYBR Green, a dye which intercalates into double stranded DNA. Specific miRNA primers, along with a Universal Primer that binds to the universal tag sequence will amplify specific miRNA sequences. 
The miScript Primer Assays are available for over a thousand human-specific miRNAs, and hundreds of murine-specific miRNAs. Relative quantification method was used here to quantify the expression of miRNAs. To correct for variability amongst different samples, expression levels of a target miRNA is normalized to the expression levels of a reference gene. The choice of a gene on which to normalize the expression of targets is critical in relative quantification method of analysis. Examples of reference genes typically used in this capacity are the small RNAs RNU6B, RNU44, and RNU48 as they are considered to be stably expressed across most samples. In this protocol, RNU6B is used as the reference gene.

MicroRNA Detection in Prostate Tumors by Quantitative Real-time PCR qPCR

Quantitative Real Time polymerase chain reaction (qPCR) is a rapid and sensitive method to investigate the expression levels of various microRNA (miRNA) molecules in tumor samples. Using this method expression of hundreds of different miRNA molecules can be amplified, quantified, and analyzed from the same cDNA template.

定量的リアルタイムPCR（定量PCR）による前立腺腫瘍におけるマイクロRNAの検出

MicroRNAs (miRNAs) are single-stranded, 18&#x2013;24 nucleotide long, non-coding RNA molecules. They are involved in virtually every cellular process ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in vivo optical and &#956;CT imaging allow the visualization of biological phenomena in an anatomical context. These imaging modalities may be especially useful to study conditions that impact bone. In particular, orthopaedic implant infections are an important problem in clinical orthopaedic surgery. These infections are difficult to treat because bacterial biofilms form on the foreign surgically implanted materials, leading to persistent inflammation, osteomyelitis and eventual osteolysis of the bone surrounding the implant, which ultimately results in implant loosening and failure. Here, a mouse model of an infected orthopaedic prosthetic implant was used that involved the surgical placement of a Kirschner-wire implant into an intramedullary canal in the femur in such a way that the end of the implant extended into the knee joint. In this model, LysEGFP mice, a mouse strain that has EGFP-fluorescent neutrophils, were employed in conjunction with a bioluminescent Staphylococcus aureus strain, which naturally emits light. The bacteria were inoculated into the knee joints of the mice prior to closing the surgical site. In vivo bioluminescent and fluorescent imaging was used to quantify the bacterial burden and neutrophil inflammatory response, respectively. In addition, &#956;CT imaging was performed on the same mice so that the 3D location of the bioluminescent and fluorescent optical signals could be co-registered with the anatomical &#956;CT images. To quantify the changes in the bone over time, the outer bone volume of the distal femurs were measured at specific time points using a semi-automated contour based segmentation process. Taken together, the combination of in vivo bioluminescent/fluorescent imaging with &#956;CT imaging may be especially useful for the noninvasive monitoring of the infection, inflammatory response and anatomical changes in bone over time.

Combined In vivo Optical and  &amp;#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined optical and &#956;CT imaging in a mouse model of orthopaedic implant infection, utilizing a bioluminescent engineered strain of Staphylococcus aureus, provided the capability to noninvasively and longitudinally monitor the dynamics of the bacterial infection, as well as the corresponding inflammatory response and anatomical changes in the bone.

合わせた<em&gt;インビボ</em&gt;光とμCTイメージングマウスにおける整形外科インプラント感染で感染症、炎症、および骨解剖学を監視する

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

Substernal Thyroid Biopsy Using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the lack of a standardized definition for STG. Biopsy is often required to differentiate benign from malignant lesions. Unlike cervical thyroid, the overlying sternum precludes ultrasound-guided percutaneous fine needle aspiration of STG. Consequently, surgical mediastinoscopy is performed in the majority of cases, causing significant procedure related morbidity and cost to healthcare. Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA) is a frequently used procedure for diagnosis and staging of non-small cell lung cancer (NSCLC). Minimally invasive needle biopsy for lesions adjacent to the airways can be performed under real-time ultrasound guidance using EBUS. Its safety and efficacy is well established with over 90% sensitivity and specificity. The ability to perform EBUS as an outpatient procedure with same-day discharges offers distinct morbidity and financial advantages over surgery. As physicians performing EBUS gained procedural expertise, they have attempted to diversify its role in the diagnosis of non-lymph node thoracic pathologies. We propose here a role for EBUS-TBNA in the diagnosis of substernal thyroid lesions, along with a step-by-step protocol for the procedure.

Substernal thyroid lesions are common, and need to be differentiated from malignancy. Obtaining percutaneous fine needle biopsy is not possible due to its retrosternal location. This article proposes a protocol for biopsy of substernal thyroid lesions using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA).

気管支超音波ガイド下経気管支穿刺吸引を使用して、胸骨下甲状腺生検

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the ...

Preparation and Respirometric Assessment of Mitochondria Isolated from Skeletal Muscle Tissue Obtained by Percutaneous Needle Biopsy

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining samples of human Vastus lateralis, isolating mitochondria from minimal amounts of skeletal muscle tissue, and plate based respirometric profiling using an extracellular flux (XF) analyzer. Comparison of respirometric profiles obtained using 1.0, 2.5 and 5.0 &#956;g of mitochondria indicate that 1.0 &#956;g is sufficient to measure respiration and that 5.0 &#956;g provides most consistent results based on comparison of standard errors. Western blot analysis of isolated mitochondria for mitochondrial marker COX IV and non-mitochondrial tissue marker GAPDH indicate that there is limited non-mitochondrial contamination using this protocol. The ability to study mitochondrial respirometry in as little as 20 mg of muscle tissue allows users to utilize individual biopsies for multiple study endpoints in clinical research projects.

Methods for biopsy of Vastus lateralis, preparation of purified mitochondria, and respirometric profiling are described. The use of small muscle volume makes this technique suitable for clinical research applications.

経皮針生検によって得られた骨格筋組織から単離されたミトコンドリアの調製と呼吸計測評価

Respirometric profiling of isolated mitochondria is commonly used to investigate electron transport chain function. We describe a method for obtaining ...

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

マウスの簡単な重要なサイズの大腿欠陥モデル

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

A Simple Device to Rapidly Prepare Whole Mounts of the Mouse Intestine

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe the use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations of superior and uniform quality to that which can be achieved by hand. The device comprises a base that holds 4 stainless steel rods and a top, which acts a cutting guide. The rods are inserted into the lumen of the small intestine [divided into thirds] and the colon. The rods and samples are then placed over a piece of filter paper or card into the holding slots in the base of the device. The top of the device is then positioned and serves as a cutting guide. The two angled sections in the center of the top piece are used to guide a knife or scalpel and cut the intestines longitudinally on the top of the rods. Once the intestinal sections have been cut, the top is removed and the card,&#160;tissue and rods gently removed from the device and placed on the bench. The rods are then gently rolled sideways to flatten and stick the intestinal segments onto the underlying piece of filter paper or card. The final preparation can then be examined or fixed and stored for later analysis. The preparations are invaluable for the study of intestinal changes in normal or genetically modified mouse models. The preparations have been used for the study and quantification of the effects of inflammation (colitis), damage, pre-cancerous lesions (aberrant crypt foci (ACFs) and mucin depleted foci (MDFs)) and polyps or tumors.

The use of a simple device to cut and &#8216;roll&#8217; mouse intestines to rapidly prepare whole mount preparations is described.

急速にマウス腸のホールマウントを準備するための簡単​​な装置

Preparing whole mounts of the mouse small intestine and colon for subsequent analysis or quantification can be time consuming and difficult. We describe ...

Generation of Prostate Cancer Patient Derived Xenograft Models from Circulating Tumor Cells

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, biologic studies, and personalized medicine strategies. Circulating tumor cells (CTC) play a critical role in tumor metastasis and have been isolated from patients with several tumor types. Recently, CTCs have been used to generate PDX experimental models of breast and prostate cancer. This manuscript details the method for the generation of prostate cancer PDX models from CTCs developed by our group. Advantages of this method over conventional PDX models include independence from surgical sample collection and generating experimental models at various disease stages. Density gradient centrifugation followed by red blood cell lysis and flow cytometry depletion of CD45 positive mononuclear cells is used to enrich CTCs from peripheral blood samples collected from patients with metastatic disease. The CTCs are then injected into immunocompromised mice; subsequently generated xenografts can be used for functional studies or harvested for molecular characterization. The primary limitation of this method is the negative selection method used for CTC enrichment. Despite this limitation, the generation of PDX models from CTCs provides a novel experimental model to be applied to prostate cancer research.

This manuscript details a method used to generate prostate cancer patient derived xenografts (PDXs) from circulating tumor cells (CTCs). The generation of PDX models from CTCs provides an alternative experimental model to study prostate cancer; the most commonly diagnosed tumor and a frequent cause of death from cancer in men.

前立腺癌患者の発生は、循環腫瘍細胞からの異種移植モデルを派生します

Patient derived xenograft (PDX) models are gaining popularity in cancer research and are used for preclinical drug evaluation, biomarker identification, ...

A Simple Mechanical Procedure to Create Limbal Stem Cell Deficiency in Mouse

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with conjunctiva. Patients suffer from recurrent corneal defects, pain, inflammation, and loss of vision.
Previously, a murine model of LSCD was described and compared to two other models. The goal was to produce a consistent mouse model of LSCD that both mimics the phenotype in humans and lasts long enough to make it possible to study the disease pathophysiology and to evaluate new treatments. Here, the technique is described in more detail.
A motorized tool with a rotating burr has been designed to remove the rust rings from the corneal surface or to smooth the pterygium bed in patients. It is a suitable device to create the desired LSCD model. It is a readily available, easy-to-use tool with a fine tip that makes it appropriate for working on small eyes, as in mice. Its application prevents unnecessary trauma to the eye and it does not result in unwanted injuries, as often is the case with chemical injury models. As opposed to a blunt scraper, it removes the epithelium with the basement membrane. In this protocol, the limbal area was abraded two times, and then the whole corneal epithelium was shaved from limbus to limbus. To avoid stroma injury, care was taken not to brush the corneal surface once the epithelium was already removed.

The following article provides an easy, reproducible technique to effectively create a sustainable mouse model of limbal stem cell deficiency (LSCD). This animal model is useful in testing and comparing the efficacy of treatments for limbal stem cell diseases.

マウスで輪部幹細胞欠損を作成するための単純な機械的手順

Limbal stem cell deficiency (LSCD) is a state of malfunction or loss of limbal epithelial stem cells, after which the corneal epithelium is replaced with ...

An Anatomical Study of Nerves at Risk During Minimally Invasive Hallux Valgus Surgery

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional orientation and localization of structures that are not directly visible to the surgeon. This is especially critical for structures at risk like nerves or blood vessels. Optimization of the handling of cadaveric material and the combination of multiple techniques compensate for the limited availability of adequate specimens. The described protocol combines anatomical plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to help localize relevant structures, such as nerves, arteries, veins and to correctly position the portals during MIS procedures. Depiction of these structures in anatomy textbooks can differ from what is encountered in the surgical field; and for this reason, new anatomical studies with a surgical orientation are needed. However, this is a complex, time-consuming technique requiring specific training. The anatomical references described with the so-called 'clock method' provide the surgeon with an easy and reproducible system to locate the path of the nerves at risk in Hallux Valgus MIS procedures. This model can be extrapolated to many other minimally invasive surgical procedures.

Minimally invasive surgical (MIS) procedures rely on anatomical references to localize structures not directly visible to the surgeon. This manuscript describes a combined method of plane-by-plane dissection and sectional anatomy of fresh-frozen specimens to locate the structures at risk during MIS procedures.

低侵襲外反母趾外反母趾手術中リスクで神経の解剖学的研究

The growing popularity of minimally invasive surgical (MIS) procedures makes it necessary that new anatomical references arise, to aid in tridimensional ...