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;

이 환자와 관련된 연구는 인간의 연구를위한 기관 심사위원회 (IRB) 승인을 받았다. 1. 환자 준비 및 위치 <ol><li> 프로 시저 제품군에 환자를 데려와 무릎이 가슴에 근육이 수축 자신의 오른쪽 또는 왼쪽에 배치합니다. </li><li> 환자, 수술 및 표시 검토 및 확정함으로써 수술 시간 종료를 수행합니다. </li><li> 표시 할 때, 마취 서비스를 통해 정맥 진정 작용을 관리 할 수​​ 있습니다. </li></ol> 2. 직장 수지 검사 (DRE)을 수행 <ol><li> 장소의 크기를 두 손 위에 적절한 장갑. </li><li> 직장 볼트로 윤활 후 장갑을 낀 손가락을 삽입하고 경직장 초음파 동안 집중 관심을 필요로하는 잠재적 의심스러운 영역에 대한 느낌 전립선을 검사합니다. (10) </li></ol> 포비돈 요오드 직장 준비 3. 창조 (PIRP) <ol><li> 10 % 15ml를 붓고멸균 분지에 시판 포비돈 요오드 용액. </li><li> 이 분지에 시중에서 판매하는 1 % 리도카인 젤의 5 ML을 추가합니다. </li><li> 포비돈 요오드 슬러리를 만들기 위해 15 초 동안 내용물을 혼합한다. </li></ol> PIRP의 4 차 관리 <ol><li> 문화 멸균 직장 면봉 직장 볼트를 사용합니다. </li><li> , 4cm의 X에게 4cm 멸균 거즈를 열고 포비돈 요오드 슬러리에 담근 후 직장 금고에 넣습니다. 만 거즈의 작은 꼬리는 항문에서 냄새가 나는데 남아 있습니다. </li><li> 이 제거되어 폐기 된 후 2 분 동안 자리에 거즈를 남겨주세요. </li></ol> PIRP의 5 차 관리 <ol><li> PIRP 솔루션 항문 주위 영역을 페인트 일회용 부인과 면봉을 사용합니다. </li><li> 항문에서 3cm의 거리에 직장 볼트로이 부인과 면봉을 삽입합니다. </li><li> 상기 PIRP 직장 점막에의 노출을 최대화하기 위해 직장 VAU 면봉유사한 방식으로, LT를 적어도 세 번. </li><li> 포비돈 요오드 용액은 2 건조하도록 허용 -. 이전의 표준 경직장 초음파 검사 및 후속 조직 검사를 진행하기에 3 ​​분 11 PIRP의 살균 효과가 노출 된 점막에 건조 솔루션을 필요로이 필수적이다. </li><li> 문화 멸균 직장 면봉 직장 볼트를 사용합니다. </li></ol> 6. 경직장 초음파 검사 및 조직 검사 <ol><li> 항문을 통해 초음파 프로브를 삽입합니다. </li><li> 모든 비뇨 관행에 부합하는 표준 방식으로 전립선 이후 조직 검사의 측정을 수행합니다. (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 PNB을 다음과 같은 감염은 매우 혈관 전립선에 직장 볼트 박테리아의 전위에 차 있습니다. 이 원리에 기초하여, 우리는 전립선 생검 이전 미생물 콜로니 수를 감소시키는 등의 방부제, 포비돈 요오드 등의 항목을 사용하는 간편한 방법을 살펴 보았다. 우리의 경험은 조직 검사의 시간이 PIRP 요법의 사용과 0.6 % (P = 0.02)에 4.3 %의 감소를 강조한다. 또한, 우리는 직장 볼트 미생물의 97 % 감소가 전?...

전립선 암은 남성에서 가장 일반적인 암 및 암 관련 사망의 두번째 주요 원인이다. 2014 년, 약 23만3천가지 경우는 거의 30,000 사람들이 암에 굴복하여 미국에서 진단됩니다. (1) 전립선 특이 항원 (PSA) 혈액 검사와 직장 수지 검사는 (DRE)이, 경직장 전립선 암 검진에 필수적인 역할을하는 동안 초음파 유도 전립선 생검 (TRUS PNB)의 조직 학적 진단을 얻을 수있는 가장 일반적인 방법이다. TRUS PNB은 환자의 항문에 초음파 프로브를 삽입하고, 일반적으로 단지 전방있는 전립선의 주연 영역의 12-14 생검을 얻는 것을 포함한다. 각각의 바늘 생검은 매우 혈관 전립선에 직장 벽을 통해 18 G 바늘의 통과를 필요로한다. 따라서, 세균의 전위에 의한 출혈과 감염의 위험이 이론적 절차에 따라 존재한다. 없음불구는 보험 수리적 합병증 후 절차는 역사적으로 낮은 유지하고있다. 이전 TRUS PNB에 항생제 예방은 일상적으로 가장 많이 사용되는 에이전트 인 퀴놀론 계 항생제로 규정되어있다. 적절한 예방에도 불구하고, 현대 조사 후 조직 검사 감염성 합병증의 증가를 연루. (2) 또한, 연구 결과에 기인 한 퀴놀론 내성 미생물, 특히 E.의 출현에 TRUS PNB을 다음과 같은 감염성 합병증이 상승 대장균. 3 퀴놀론의 저항이 상승하는 속도는 TRUS PNB 절차 예방 대체 요법의 필요성을 시사한다. 이를 위해 여러 가지 방법을 조사 중에있다. 한 가지 방법은 경구 용 퀴놀론와 함께 조직 검사시 정맥 또는 근육 항생제의 관리를 포함한다. (4)이 전략은 TRUS PNB 다음 패혈증 이벤트를 줄일 수 있지만,이 접근법의 주요 한계는 더욱 가족이 항생제 내성 미생물을 개발하기위한 전위이다. 또, 최근 연구 방법론 5. 퀴놀론 - 내성 직장 식물 집락 환자를 선별하기 전 생검 직장 면봉의 사용을 포함 이러한 생물체가 검출되면, &quot;타겟&quot;항생제 요법은 식별 내성 세균의 감도 프로파일에 기초하여 사용될 수있다 변형. 방법론은 우아하지만, 선택적으로, 이러한 면봉을 획득 퀴놀론 선택 배지에 배양 및 항생제를 재봉의 실제 과정은 이후 많은 임상 사례에서 부족 될 수있는 임상 및 실험실 인프라가 필요합니다. TRUS PNB 감염을 제한하는 대안 전략을 제시 할 수있다 조직 검사 이전에 직장 금고 미생물의 집락 수를 줄이기 위해 살균 주제의 관리. 포비돈의 iodone은 저렴하고 쉽게 사용할 수 에이전트수술 사이트에 적용 할 때 감소 박테리아 수를 기록. 모두 대장 암 및 부인과 수술의 응용 프로그램은 잘 알려져있다. 따라서, 직장 제제로서 povidine 요오드의 사용은 종래 생검 추가적인 준비 없이도 감소 TRUS PNB 감염에, 제조 가능한 간단하고 경제적 인 방법을 제시한다. 비뇨 문학에 대해, 이전에 작업 TRUS PNB 다음과 같은 임상 적 감염의 감소를 지적했다. 6-9이 동영상은 PIRP의 기술은 임상 관행에 통합 용이성과 단순성을 강조 보여줍니다. TRUS PNB을받은 모든 환자는 초기 전립선 암 검진에 대한 추천 또는 초기 평가에 대한 전문 비뇨기과 클리닉에서 볼 수 있었다. 이 평가는 환자의 혈청 전립선 특이 항원 (PSA)와 직장 수지 검사 (DRE)의 성능 검토를 수반. PSA 나 DRE 하나의 이상이의 추천 메시지TRUS PNB. TRUS PNB 절차는 모든 환자와 감염, 출혈을 포함하여 관련 위험 논의하고, 요폐는 명확했다. 모든 환자는 퀴놀론 계 약물 (예, 시프로플록사신) 또는 트리 메소 프림 - 설파 (Bactrim) 조직 검사 이전에 하나 경구 항생제 예방 삼일을 받았다.

<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

단일 기관과 인구 기반 연구는 경직장 초음파 유도 전립선 생검 (TRUS PNB) 다음과 같은 감염성 합병증이 증가하고 있음을 강조. 이러한 감염은 직장 볼트를 식민지화하고 조직 검사 절차 중에 혈류로 전좌되어 내성 미생물을 퀴놀론에 크게 기인한다. 생검시 포비돈 요오드 직장 제제 (PIRP)는 TRUS PNB 따라 직장 균 집락 따라서 얻어진 감염을 줄이기위한 간단하고 재현 가능한 방법이다. 모든 환자는 이전에 조직 검사에 경구 항생제 치료의 삼일을 투여한다. PIRP 기술은 초기 TRUS PNB위한 표준 방식으로 환자를 위치시키는 것을 포함한다. 디지털 직장 검사 후, 시판 포비돈 요오드의 10 % 용액의 15 mL의 슬러리를 만들기 위해 1 % 리도카인 젤리 5 ㎖와 혼합된다. 4cm X 4cm 멸균 거즈는 SOA입니다이 슬러리에 KED 후 제거한 후 2 분 동안 직장 볼트 삽입. 그 후, 일회용면 부인과 면봉은 항문 주위 영역과 항문에서 3cm의 거리에 직장 금고를 모두 칠하는 데 사용됩니다. 이전의 표준 경직장 초음파 검사 및 후속 조직 검사를 진행하기로 3 분 - 포비돈 요오드 용액을 2 건조 할 수있다. 이 PIRP 기술은 0.6 % (P = 0.02)에 4.3 %에서 후 조직 검사 감염의 연관 감소 2012 년 3 월 이후 우리의 기관에서 연습하고있다. 이 예방 요법의 주된 장점은 감염을 감소시키기 쉽게 사용할 저렴 에이전트의 사용과 단순성 및 재현성이다. 또한, 기술은 다 약제 내성균의 전위가 전파와 추가 환자 전신 항생제 노출 피한다. TRUS PNB에서 PIRP의 사용은, 그러나, 요오드 또는 조개 환자에는 적용되지 않습니다llergies.

이 PIRP 기술은 165 명의 환자가이 품질 개선 조치에 등록 된과 (현재까지) 2012 년 월부터 연습에왔다. 이 시간에, 단 하나의 환자 (0.6 %), 전신 패혈증의 형태로 감염 합병증을 경험했다. 이 환자는 다 약제 내성 E. 집락시켰다 대장균 균주. 이 환자 PIRP의 관리에 따라 직장 면봉은 직장 금고는 여전히 2.1 × 105 CFU / ml의 세균이 이에 PIRP 적절하게 직장 볼트를 처리 여부 질문과 식민?...

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.

전립선 바늘 생검시 포비돈 요오드 직장 준비 절차 감염의 위험을 줄이기 위해 간단하고 재현 수단이다

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 조회수.  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.

비디오: 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.

Intravital 이미징을 사용하여 실시간으로 종양의 Nanoparticle의 이해 평가

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

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의학

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.

정량 실시간 PCR (qPCR)에 의한 전립선 종양에 MicroRNA 감지

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.

결합<em&gt; 생체</em&gt; 광학 및 μCT 이미징 마우스의 정형 외과 임플란트 감염에 감염, 염증 및 뼈 해부학을 모니터링

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).

흉골 하 갑상선 조직 검사는 기관지 초음파 유도 기관지 바늘 흡인을 사용하여

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.

경피적 바늘 생검에 의해 획득 골격근 조직에서 고립 된 미토콘드리아의 준비 및 호흡계 평가

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.

마우스의 간단한 임계 크기의 대퇴 결함 모델

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.

간단한 장치는 빠르게 마우스 소장의 전체 마운트를 준비하는 방법

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.

전립선 암 환자의 발생은 종양 세포를 순환에서 이종 이식 모델을 파생

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.

간단한 기계 절차 마우스에서 윤부 줄기 세포 결핍을 만드는 방법

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.

최소 침 습 Hallux Valgus 수술 중 위험에 신경의 해부학 적 연구

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